SYNERGISTIC COMPOSITIONS OF SOLUBLE COMPLEMENT RECEPTORS AND COMPOUNDS THAT INHIBIT COMPLEMENT AND/OR SUPPRESS IMMUNE ACTIVITY

Abstract: The present invention relates to synergistic therapeutic compositions of synergistic combinations of organic and biological compounds which inhibit complement and/or possess immunosuppressive activity, and the therapeutic uses thereof. The organic compounds of the invention are compounds which exhibit immunosuppressive and/or anti-inflammatory activity. Such organic compounds include but are not limited to K-76, K-76 COOH and derivatives and synthetic analogs thereof, antimetabolites, antimitotic drugs, cyclosporine, steroids, and non-steroidal anti-inflammatory agents. In a particular aspect, the organic compounds of the invention include substituted dihydrobenzofurans, substituted and unsubstituted spirobenzofuran-2(3H)-cycloalkanes, and their open chain intermediates which exhibit such inhibitory ativities. The biological compounds of the invention are complement receptors or fragments thereof and soluble members of the complement receptor family that contain the conserved SCR motif and that are able to inhibit complement activity. The biological compounds exhibit at least one of the functions associated with complement receptors and/or SCR containing proteins. The invention relates to the use of mixtures of these compounds for therapy of immune and/or inflammatory disorders.

Description: SYNERGISTIC COMPOSITIONS OF SOLUBLE COMPLEMENT RECEPTORS AND COMPOUNDS THAT INHIBIT COMPLEMENT AND/OR SUPPRESS IMMUNE ACTIVITY 1. FIELD OF THE INVENTION The present invention relates to therapeutic composi¬ tions of organic and biological compounds, which synergis- tically or additively inhibit complement and/or possess immunosuppressive activity, and therapeutic uses thereof. The organic compounds of the invention are compounds which exhibit complement inhibitory immunosuppressive, and/or anti-inflammatory activity. The biological compounds exhibit at least one of the functions associated with complement receptors and/or SCR- containing proteins. The invention relates to the use of mixtures of these com- pounds for therapy of immune and/or inflammatory disor¬ ders. 2. BACKGROUND OF THE INVENTION 2.1. THE COMPLEMENT SYSTEM The complement system is a group of proteins that constitutes about 10 percent of the globulins in the normal serum of humans (Hood, L.E. et al. 1984, Immunolo¬ gy, 2d Edition, The Benjamin/Cummings Publishing Co., Menlo Park, California, p. 339) . Complement (C) plays an i.mportant role i.n the medi.ati.on of i.mmune and allergic reactions (Rapp, H.J. and Borsos, T., 1970, Molecular Basis of Complement Action, Appleton-Century-Crofts (Meredith) , New York) . The activation of C components leads to the generation of a group of factors, including . . . . . chemotactic peptides that mediate the inflammation associ¬ ated with complement-dependent diseases. The sequential activation of the complement cascade may occur via the classical pathway involving antigen-antibody complexes, or by an alternative pathway which involves the recognition of certain cell wall polysaccharides. The activities mediated by activated complement proteins include lysis of target cells, chemotaxis, opsonization, stimulation of vascular and other smooth muscle cells, degranulation of 5 mast cells, increased permeability of small blood vessels, directed migration of leukocytes, and activation of B lymphocytes, macrophages and neutrophils (Eisen, H.N., 1974, Immunology, Harper & Row, Publishers, Inc., Hagerstown, Maryland, p. 512) . . Q During proteolytic cascade steps, biologically active peptide fragments, the anaphylatoxins C3a, C4a, and C5a (See WHO Scientific Group, WHO Tech. Rep. Ser. 1977, 606, 5 and ref rences cited therein) , are released rom the third (C3) , fourth (C4) , and fifth (C5) native complement 15 components (Hugli, T.E. CRC Crit. Rev. Immunol. 1981, l r 321; Bult, H. and Herman, A.G. Agents Actions 1983, 13, 405) . The C5a fragment, a cationic peptide derived from the first 74 amino acids of the amino-terminus of the C5 alpha subunit (Tack, B.F. et al. Biochemistry 1979, 18, 201490) , is of particular pathological relevance. Regula¬ tion of C5a activity is by the endogenous plasma enzyme carboxypeptidase N (E.C. 3.4.12.7), which rapidly removes the carboxy-terminal arginine from C5a, producing the less potent but still active C5a des Arg. Reported effects of 5 C3a and C5a upon specific immune responses are listed in Table I. 0 5 TABLE I EFFECTS OF COMPLEMENT COMPONENTS C3a AND C5a ON SPECIFIC IMMUNE RESPONSES Immune Response C3a C5a/C5a des Arcr Specific antibody production in response to sheep red blood cells Suppression Enhancement Polyclonal antibody production in response to Fc antibody fragment Suppression Enhancement T cell proliferation in response to tetanous toxoid Suppression Enhancement T cell proliferation in mixed lymphocyte reaction No effect Enhancement T cell-mediated cytotoxicity Suppression Enhancement Among the wide variety of biological activities exhibited by C5a are contraction of smooth muscle (Wissler, J.H. Eur. J. Immunol. 1972, 2, 73), degranulation of mast cells (Johnson, A.R. et al., Immunol. 1975, 28, 1067), secretion of azurophilic granular enzymes from polymorphonuclear neutrophils (PMN) (Webster, R.O. et al., Immunopharmacol. 1980, 2, 201), and the chemotaxis of PMN (Wissler, J.H. Eur. J. Immunol. 1972, 2, 73; Becker, E.L. Trends Pharmacol. Sci. 1983, 4, 223) (Table II). TABLE II BIOLOGICAL EFFECTS OF C5a 5 I. Stimulation of neutrophil functions involved in inflammation A. chemotaxis B. chemokinesis 10 C. aggregation D. lysoso al enzyme release E. generation of toxic oxygen products II. Smooth muscle effects A. stomach smooth muscle contraction 15 B. vasodilation II. Promotion of histamine release A. mast cells B. basophils IV. Immunoregulatory effects 20 The active chemotactic factor in vivo is considered to be C5a des Arg (Becker, E.L. Trends Pharmacol. Sci. 1983, 4, 223) . The C5a or C5a des Arg fragments have been implicated 25 in the infiltration of PMN (the chemotacti.c effect) i.n rheumatoid arthritis, certain forms of glomerulonephritis, experimental vasculitides such as the Arthus reaction, the acute pneumonitis produced by the instillation of chemo¬ tactic factors into the lungs of experimental animals with 30 resulting release of leukotπenes C-4 and D-4 (LTC 4 and LTD 4 ) , etc. In addition, the interactions between C5a and neutrophils have been considered to underlie tissue damage in several clinical situations. For instance, there «- 5 exists a growing body of evidence for the role of oxygen-derived free radicals in mediating myocardial tissue injury during myocardial ischemia and, in particu¬ lar, during the phase of myocardial reoxygenation and reperfusion. Among a number of possible sources of these c radicals, the polymorphonuclear neutrophil has been the focus of primary attention. Studies have documented that neutrophil depletion or suppression of neutrophil function results in a significant salvage of myocardial tissue that is subjected to a period of regional ischemia followed by 10 reperfusion (Simpson, P.J. and Lucchesi, B.R. J. Lab. Clin. Med. 1987, 110(1), 13-30). Neutrophil depletion in dogs resulted in significantly smaller myocardial infarcts after 90 minute occlusion with 24 hour reperfusion (Jolly, S.R. et al. Am. Heart J. 1986, 112, 682-690). 15 2.2. ORGANIC COMPOUNDS WHICH INHIBIT COMPLEMENT, AMELIORATE INFLAMMATION, AND/OR POSSESS IMMUNOSUPPRESSIVE ACTIVITY Many chemicals have been reported to diminish comple¬ ment-mediated activity. Such compounds include: amino 20 acids (Takada, Y. et al. Immunology 1978, 34, 509); phosphonate esters (Becker, L. Biochem. Biophy. Acta 1967, 147, 289); polyanionic substances (Conrow, R.B. et al. J. Med. Chem. 1980, 23, 242); sulfonyl fluorides (Hansch, C. ; Yoshimoto, M. J. Med. Chem. 1974, 17, 1160, and references 25 cited therein); polynucleotides (DeClercq, P.F. et al. Biochem. Biophys. Res. Commun. 1975, 67, 255); pimaric acids (Glovsky, M.M. et al. J. Immunol. 1969, 102, 1) ; porphines (Lapidus, M. and Tomasco, J. Immunopharmacol. 1981, 3, 137); several antiinflammatories (Burge, J.J. et 30 al. J. Immunol. 1978, 120, 1625); phenols (Muller-Eberhard, H.J. 1978, in Molecular Basis of Biolog¬ ical Degradative Processes, Berlin, R.D. et al., eds. Academic Press, New York, p. 65) ; and benzamidines (Vogt, __ W. et al Immunology 1979, 36, 138). Some of these agents 35 express their activity by general inhibition of proteases and esterases. Others are not specific to any particular intermediate step in the complement pathway, but, rather, inhibit more than one step of complement activation. Examples of the latter compounds include the benza idines, which block Cl, C4 and C5 utilization (Vogt, W. et al. Immunol. 1979, 36, 138). Many organic compounds that possess immunosuppressive activities and act to ameliorate inflammation are also well known in the art. 2.3. SPIROBENZOFURAN-2f3H.-CYCLOALKANES AND K-76 K-76 is a fungal metabolite from Stachvbotrys complementi nov. sp. K-76. Metabolite K-76 has a drimane skeleton combined with a benzene ring attached through a spirofuran, and has been determined as 6,7-diformyl- 3' ,4 « , a• ,5' ,6 l ,7• ,8' ,8 '-octahydro-4,6' ,7•-trihydroxy- 2*,5 « ,5• ,8 •-tetramethyl spiro [1• (2 ■ H)-naphthalene-2(3H)- benzofuran] (Kaise, H. et al. J. Chem. Soc. Chem. Commun. 1979, 726). The monocarboxylic acid derivative, K-76 COOH, is obtained when K-76 is selectively oxidized by silver oxide (Corey, E.J. and Das, J. J. Amer. Chem. Soc. 1982, 104, 5551). Both K-76 and K-76 COOH have been shown to inhibit complement mainly at the C5 step (Hong, K. et al. J. Immunol. 1979, 122, 2418; Miyazaki, W. et al. Microbiol. Immunol. 1980, 24, 1091) . In a classical hemolytic reac¬ tion system, hemolysis of sensitized sheep erythrocytes by guinea pig serum was reduced 50% by K-76 at 7.45 x 10 "5 M, or K-76 COONa at 3.41 x 10- 4 M (Hong, K. et al. J. Immunol. 1979, 122, 2418; Miyazaki, W. et al. Microbiol. Immunol. 1980, 24, 1091) . Similar results were observed in a hemolytic reaction system via the alternative pathway of complement activation. Both K-76 and K-76 COOH prevented the generation of a chemotactic factor from normal human complement (Bumpers, H. and Baum, J. J. Lab. Clinc. Med. 1983, 102, 421). K-76 has been shown to reduce the amount of protein excreted in urine of rats with nephrotoxic-glomerulonephritis (Iida, H. , et al., Clin. Exp. Immunol. 1987, 67, 130-134), and is 5 reported to greatly increase the survival of mice with a spontaneous systemic lupus erythematosis-like disease and to suppress Forssman shock in guinea pigs and mice (Miyazaki, W. et al. Microbiol. Immunol. 1980, 24, 1091) . At high concentrations of K-76 or K-76 COOH, some inhibi- 0 tion of the reactions of C2, C3, C6, C7, and C9 with their respective preceding intermediaries is exhibited. Howev¬ er, both compounds 1 inhibitory action is mainly the gener¬ ation in vitro of EAC1, 4b,2a,3b,5b (sensitized sheep erythrocytes carrying the indicated complement components) 5 from C5 and EACl,4b,2a,3b; the acceleration of the decay of any EACl,4b,2a,3b,5b present; and blocking generation of the chemotactic peptides (Hong, K. et al. J. Immunol. 1981, 127, 109; Ramm, L.E., et al. Mol. Immunol. 1983, 20, 155) . 0 K-76 COOH is also reported to be an anti-hepatitic agent (West German Patent Application, Publication No. 3,031,788, published March 12, 1981, by Shinohara, M. et al.), and possesses the ability to inhibit antibody-depen¬ dent cell-mediated cytotoxicity and natural killer lytic 5 activity (Hudig, D. et al. J. Immunol. 1984, 133, 408-413). K-76 or K-76 COOH has also been reported to inhibit the C3b inactivator system of complement (Hong, K. et al. J. Immunol. 1981, 127, 104-108). Semi-synthetic derivatives of K-76 have been patented as anti-allergy, ° anti-tumor, and anti-nephritic agents (Belgium Patent No. 867,095, published November 16, 1978, by Shinohara, M. et al.). The isolation of K-76, its uses in the treatment of autoimmune diseases, and the preparation of its deriva¬ tives have been described in a number of patents (See 5 Japanese Patent Appli.cati.ons (Kokai.) , Publi.cati.on Nos. 54 092680 (published July 23, 1979) 54 106458 (published August 21, 1979) 57 083281 (published May 25, 1982), by Shinohara, M. et al. ; Japanese Patent (Kokoku) No. 85 030289 (published March 20, 1979)). ς A number of additional compounds which contain the substructure of a spirobenzofuran-2(3H)-cycloalkane are known. These compounds include griseofulvin (Weinberg, E.D., 1981, in Principles of Medicinal Chemistry, 2d Ed., Foye, W.O., ed. , Lea & Febiger, Philadelphia, PA., p. 10 813) , isopannarin (Djura, P. and Sargent, M.V. Aust. J. Chem. 1983, 36, 1057), and metabolites of Siphonodictvon cora11i-phacrum (Sullivan, B., et al. Tetrahedron 1981, 37, 979) . 15 2.4. COMPLEMENT RECEPTORS C3b/C4b COMPLEMENT RECEPTOR (CR1) . The human C3b/C4b receptor, termed complement receptor I (CR1, also CD35) , is present on erythrocytes, monocytes/macrophages, granul- ocytes, B cells, some T cells, splenic follicular dendrit- 0 ic cells, and glomerular podocytes (Fearon D.T., 1980, J. Exp. Med. 152:20, Wilson, J.G. et al., 1983, J. Immunol. 131:684; Reynes, M. , et al. , 1976 N. Engl. J. Med. 295:10; Kazatchkine, M.D., et al. , 1982, Clin. Immunol. Immuno- pathol. 27:210). CR1 specifically binds C3b, C4b and 5 iC3b. CR1 can also inhibit the classical and alternative pathway C3/C5 convertases and act as a cofactor for the cleavage of C3b and C4b by factor I, indicating that CR1 also has complement regulatory functions in addition to 0 serving as a receptor (Fearon, D.T., 1979, Proc. Natl. Acad. Sci. U.S.A. 76:5867; Iida, K. and Nussenzweig, V., 1981, J. Exp. Med. 153:1138). CR1 has been shown to have homology to complement receptor type 2 (CR2) (Weis, J.J. , et al. , 1986, Proc. Natl. Acad. Sci. U.S.A. 83:5639-5643). Four allotypic forms of CR1 have been found, differ¬ ing by increments of 40,000-50,000 daltons molecular weight. All Four CR1 allotypes have C3b-binding activity (Dykman, T.R., et al., 1983, Proc. Natl. Acad. Sci. U.S.A. 5 80:1698; Wong, W.W. , et al., 1983, J. Clin. Invest. 72:685; Dykman, 20 T.R. , et al., 1984, J. Exp. Med. 159:691; Dykman, T.R. , et al., 1985, J. Immunol. 134:1787) . CR2. Complement receptor type 2 (CR2, CD21) is a ,« transmembrane phosphoprotein consisting of an extracel¬ lular domain which is comprised of 15 or 16 SCR's, a 24 amino acid transmembrane region, and a 34 amino acid cytoplasmic domain (Moore, et al., 1987, Proc. Natl. Acad. Sci. U.S.A. 84:9194-9198; Weis, et al., 1988, J. Exp. Med. 15167:1047-1066 which are incorporated herein by reference). Electron microscopic studies of soluble recombinant CR2 have shown that, like CR1, it is an extended, highly flexible molecule with an estimated contour length of 39.6 nanometers by 3.2 nanometers, in which each SCR appears as 0 a ringlet 2.4 nanometers in length (Moore, et al., 1989, J. Biol. Chem. 34:20576-20582). A form of recombinant soluble CR2 has been produced (Moore, et al., 1989, J. Biol. Chem. 264:20576-20582). In analogy to the soluble CR1 system, soluble CR2 was pro- 5 duced in a recombinant system from an expression vector containing the entire extracellular domain of the recep¬ tor, but without the transmembrane and cytoplasmic do¬ mains. This recombinant CR2 is reported to bind to C3dg in a 1:1 complex with Kd (disassociation constant) equal 0 to 27.5 mM and to bind to the Epstein-Barr proteins gp350/220 in a 1:1 complex with Kd=3.2 nM (Moore, et al., 1989, J. Viol. Chem. 264:2057620582). CR3. A third complement receptor, CR3, also binds iC3b. Binding of iC3b to CR3 promotes the adherence of 5 neutrophils to complement-acti.vating endothelial cells during inflammation (Marks, et al. , 1989, Nature 339:314). CR3 is also involved in phagocytosis, where particles coated with iC3b are engulfed by neutrophils or by acro- phages (Wright, et al. , 1982, J. Exp. Med. 156:1149; Wright, et al. , 1983, J. Exp. Med. 158:1338). CR4. CR4 (CD11) also appears to be involved in leukocyte adhesion (Kishimoto et al. , 1989, Adv. Immunol. 46:149-82) . Short Consensus Repeat (SCR) Motif. CR1 has been extensively studied, and a structural motif of 60-70 amino acids, termed the short consensus repeat (SCR) has been found. The SCR motif is tandemly repeated 30 times in the F-allotype of CR1 and additional repeat cycles occur in other allotypes. The consensus sequence of the SCR in- eludes 4 cysteines, a glycine and a tryptophan that are invariant among all SCR. Sixteen other positions are conserved, with the same amino acid or a conservative replacement being found in over half of the other 30 SCRs (Klickstein, et al. , 1987, J. Exp. Med. 165:1095-1112; Klickstein et al, 1988, J. Exp. Med., 168:1699-1717; Hourcade et al., 1988, J. Exp. Med. 168:1255-1270). The dimensions of each SCR are estimated to be approximately 2.5-3.0 nm x 2 nm x 2 nm. 2.5. COMPLEMENT MEDIATED DISEASES AND DISORDERS MANIFESTING INAPPROPRIATE COMPLEMENT ACTIVATION The complement activation pathways play a fundamental role in many human diseases and disorders. Some of the clinical implications of C5a release are listed in Table HI- Such disease conditions include but are not limited to immune complex diseases (such as systemic lupus erythe- atosus, arthritis, glomerulonephritis, and AIDS) . TABLE III Diseases and Disorders involving Complement Autoimmune Diseases Rheumatoid arthritis Immune-complex-induced vasculitis Arthritis Type II collagen-induced arthritis Acute Immunological Arthritis Myasthenia gravis Acute Gouty Arthritis Systemic lupus erythematosis Hemolytic anemia Multiple sclerosis G1omeru1oneph i is Experimental allergic neuritis Immune Complex Disorders Pulmonary Disorders Adult Respiratory Distress Syndrome Pulmonary Dysfunction - Hemodialysis Chronic Progressive Pulmonary Dis-Cystic Fibrosis Byssinosis Asbestos-Induced Inflammation Inflammation of Glomerulonephritis Inflammation of Crohn's disease Purtscher•s Retinopathy Hemorrhagic Pancreatitis Renal Cortical Necrosis Primary Biliary Cirrhosis Inflammation Nephropathology Cranial Nerve Damage in Meningitis Tumor Cell Metastasis Extended Tissue Destruction in Myocardial Infarction Extended Tissue Destructi.on i.n Burns Infectious Diseases Induced by virus Epstein-Barr Virus Associated Diseases Sjogren's Syndrome, Rheumatoid arthritis, urkitt's Lymphoma, Hodgkins Disease, virus AIDS 5 or EBV) associated B cell lymphoma, Chronic Fatigue Syndrome, parasitic diseases states (such as viral infection following allograft transplantation or AIDS) . HTLV-III / LAV / HIV (AIDS) 10 Myocardial Infarction. Recombinant tissue plasmino- gen activator (r-TPA) , which in recent clinical trials has been found to be an effective thrombolytic agent in pa¬ tients with acute myocardial infarction, was shown to 15 activate complement. A striking increase in the level of C4a, C3a, and C5a was found in patients receiving r-TPA as compared to the level of these complement peptides before administration of the drug (Bennett, W.R. et al. J.Am. Coll. Cardiol. 1987, 10(3), 627-632). Schafer and * _>π u co-workers were able to posi.tively i.dentify the depositi.on of terminal C5b-9 complement complex in myocardial cells located within zones of infarction in human tissue (J. Immunol. 1986, 137(6), 1945-1949). Likewise, the selec¬ tive accumulation of the first component of complement and 25 leukocytes in ischemic canine heart muscle has been found (Rosen, R.D. et al. Circ. Research, 1985, 57, 119-230). In one study, the depletion of complement was found to increase the blood flow in ischemic canine myocardium. This increased blood flow was found, in turn, to increase 30 the supply and utilization of oxygen in complement deplet¬ ed animals versus control animals (Grover, G.J. and Weiss, H.R. Basic Res. Cardio. 1987, 82(1}_, 57-65). A recombinant soluble form of CR1 has been produced which is able to „Jo inhibit activation of both the classical and alternative pathways in vitro. It also can suppress complement acti¬ vation in vivo and is effective in animal models of in¬ flammation and reperfusion injury associated with myocar¬ dial infarction (International Patent Application number PCT/US89/01358, published October 5, 1989 as W089/09220 and entitled "The Human C3b/C4b Receptor (CRl)"; Weissman et al., 1990, Science 249:146-151). SLE. Diminished expression of CRl on erythrocytes of patients with systemic lupus erythematosus (SLE) has been reported by investigators from several geographic regions, including Japan (Miyakawa et al., 1981, Lancet 2:493-497; Minota et al., 1984, Arthr. Rheum. 27:1329-1335), the United States (Iida et al., 1982, J. Exp. Med. 155:1427- 1438; Wilson et al., 1982, N. Engl. J. Med. 307:981-986) and Europe (Walport et al., 1985, Clin. Exp. Immunol. 59:547; Jouvin et al., 1986, Complement 3:88-96; Holme et al., 1986, Clin. Exp. Immunol. 63:41-48). Autoimmune Disease. Activation of the complement system is suggested to cause tissue injury in animal models of autoimmune diseases such as immune-complex- induced vasculitis (Cochrane, 1984, Springer Seminar Immunopathol. 7:263), glomerulonephritis (Couser et al., 1985, Kidney Inst. 29:879), hemolytic anemia (Schreiber & Frank, 1972, J. Clin. Invest. 51:575), yasthenia gravis (Lennon et al., 1978,, J. Exp. Med. 147:973; Biesecer & Gomez, 1989, J. Immunol. 142:2654), type II collagen- induced arthritis (Watson & Townes, 1985, J. Exp. Med. 162:1878), and experimental allergic neuritis (Feasby et al., 1987, Brain Res. 419:97). Immune Complex Disorders. Immune complexes are found in many pathological states including but not limited to autoimmune diseases such as rheumatoid arthritis or SLE, hematologic malignancies such as AIDS (Tayler et al., 1983, Arthritis. Rheum. 26:736-44; Inada et al., 1986, AIDS Research 2:235-247) and disorders involving autoanti- bodies and/or complement activation (Ross et al., 1985, J. Immunol 135: 2005-14). Erythrocytes are involved in the removal of circulating immune complexes via adherence to erythrocyte-CRl and may function to inhibit deposition of immune complexes in body tissue. A method of treatment for the removal and immobilization of circulating immune complexes involves the transfusion of packed erythrocytes with high CRl activity; this process is dependent upon complement consumption. AIDS. Erythrocyte CRl has a functional role in the removal of circulating immune complexes in autoimmune pa¬ tients and may thereby inhibit the deposition of immune complexes within body tissue constituents (Inada et al., 1989, Ann. Rheum. Dis. 4:287). In the development of AIDS, decremental loss of CRl activity progresses from asymptomatic seropositive homosexual volunteers to the prodromal spectrum of ARC and finally progressing to a total disappearance in overt AIDS (Inada et al. , 1986, AIDS Res. 2:235) These studies demonstrated an associa- tion of clinical disease state with a decreased CRl activ¬ ity rather than with levels of circulating immune complex. The relative loss of CRl from erythrocytes that has been observed in patients with Human Immunodeficiency Virus (HIV) infections (Tausk, F.A. , et al. , 1986, J. Clin. Invest. 78:977-982) has also been observed with leproma- tous leprosy (Tausk, F.A. , et al. , 1985, J. Invest. Der at. 85:58s-61s) . Inflammation Disorders. Complement activation has also been associated with disease states involving infla - mation. The intestinal inflammation of Crohn's disease is characterized by the lymphoid infiltration of mononuclear and polymorphonuclear leukocytes. It was found recently (Ahrenstedt et al, 1990, New Eng. J. Med. 322:1345-9) that the complement C4 concentration in the jejunal fluid of Crohn's disease patients increase compared to normal controls. Other disease states implicating the complement system in inflammation include thermal injury (burns, frostbite) (Gelfand et al., 1982, J. Clin. Invest. 70:1170; Demling et al., 1989, Surgery 106:52-9), hemodi- 5 alysis (Deppisch et al., 1990, Kidney Inst. 37:69-706; Kojima et al., 1989 Nippon Jenzo Gakkai Shi 31:91-7), and post-pump syndrome in cardiopulmonary bypass (Chenoweth et al., 1981, Complement Inflamm. 3:152-165; Chenoweth et al., 1986, Complement 3:152-165; Salama et al., 1988, N. Q Engl. J. Med. 318:408-14). Hemodialysis. The use of a hemodialyzer activates the alternative pathway. Levels of Bb, iC3b, C3a and C5a increase, but C4d levels do not change (Oppermann et al., 1988, Klin. Wochen. 66:857-864; Kojima et al., 1989; 5 Nippon Jenzo Gakkai Shi 31:91-97; Ueda et al., 1990, Nippon Jenzo Gakkai Shi 32:19-24). Terminal complement complex was demonstrated in the plasma (Kojima et al., 1989, supra) . and on the membranes of granulocytes during dialysis (Deppisch et al., 1990, Kidney. Int. 37:696-706). 0 Cardiopulmonary Bypass. The use of pump-oxygenator systems in cardiopulmonary bypass and hemodialysis has been associated with a systemic inflammation reaction, which may result in profound organ dysfunction. These effects have been collectively termed as the post-pump syndrome or post-perfusion syndrome. The clinical find¬ ings of this syndrome are similar to the biological activ¬ ities of C3a and C5a. Indeed, elevated C3a has been demonstrated in patients undergoing prolonged extracorp- oreal circulation (Chenoweth et al., 1981, Complement inflamm. 3:152-165). Increased plasma levels of SC5b-9 (Dalmasso et al., 1981, Compliment Inflamm. 6:36-48), and terminal C5b-9 complex deposits on erythrocytes and poly¬ morphonuclear cells (Salama et al., 1988, N. Engl. J. Med. 318:408-414) have been observed in these patients. One study documented the activation of complement and genera- tion of oxygen-derived free radicals during cardiopulmo¬ nary bypass. The administration of protamine during cardiopulmonary bypass further activated complement (Cavarocchi, N.C. et al. , 1986, Circulation, 74, 130-133; Kirklen, J.K. et al., 1983, J. Thorac. Cardiovasc. Surg. 86, 845-857) . Three experimental models for cardiopulmonary bypass were identified, rat (Alexander & Alani, 1983, J. Surg. Res.35:28-34) , pig (Nilsson et al., 1990, Artif. Organs 14:46-48) and cynomolgus monkey (Maeo, 1989, Nippon Kyobu Geka Gakkai Zasshi 37:2166-2174). Thermal Injury (burn) . Most of the pathologic pro¬ cesses initiated by burns are inflammatory in nature. The main complications are shock, pulmonary edema, and infec- tion. It has been shown that massive activation of the alternative complement pathway, but not the classical pathway, was observed in a model of burn injury in mice; cobra venom factor pretreatment reduced burn mortality (Gelfand et al., 1982, J. Clin. Invest. 70: 1170-1176). Elevations of plasma C3a des Arg and C4a des Arg were also detected in burn patients (Davis et al. , 1987, Surgery 102:477-484) . Adult Respiratory Distress syndrome (ARDS) . Both complement and leukocytes are reported to be implicated in the pathogenesis of adult respiratory distress syndrome (Zilow et al., 1990, Clin. Exp. Immunol. 79:151-57; Lang- lois et al., 1989, Heart Lung 18:71-84). This syndrome, also known as adult respiratory failure, shock lung, diffuse alveolar damage, or traumatic wet lungs, is char- acterized clinically by the rapid onset of severe life- threatening respiratory insufficiency that is refractory to oxygen therapy (Miescher, P.A. and Muller-Eberhard, H.J., eds., 1976, Text Book of Immunopathology, 2d Ed., Vols. I and II, Grune and Stratton, New York; Sandberg, A.L., 1981, in Cellular Functions in Immunity and Inflam- mation, Oppenheim, J.J. et al., eds., Elsevier/North Holland, New York, p. 373; Conrow, R.B., et al., 1980, J. Med. Chem 23: 242; Regal, J.F.; and Pickering, R.H., 1983, Int. J. Immunopharmacol. 104: 617). 5 Sepsis. The mortality of sepsis is high, mainly due to complications such as shock and ARDS. Activation of the complement system via the classical pathway is sug¬ gested to be involved in the development of fatal compli¬ cations in sepsis (Hack et al., 1989, Am. J. Med. 86:20- 10 26 >- Barotrauma. It has been hypothesized that the phe¬ nomena of decompression sickness (DCS) are mediated by complement. The complement system was activated in rab¬ bits with CDS. When these rabbits were pharmacologically 1 5 decomplemented in vivo. they did not develop DCS (Ward et al., 1990, Undersea Bio ed. Res. 17:51-66). Allograft Rejection. The complement system is also involved in hyperacute allograft and hyperacute xenograft rejection (Knechtle et al., 1985, J. Heart Transplant θ4(5):541; Guttman, 1974, Transplantation 17:383; Adachi et al., 1987, Trans. Proc. 19(1):1145; Pruitt and Bollinger, 1991, J. Surg. Res. 50:350-355; Pruitt et al., 1991, Transplantation 52:868-873; Migagowa et al., 1988, Transplantation 46:825). 5 Interleukin-2 Therapy. Complement activation during immunotherapy with recombinant IL-2 appears to cause the severe toxicity and side effects observed from IL-2 treat¬ ment (Thijs et al., 1990, J. Immunol. 144:2419). CR2 Implicated Diseases. Such disease conditions in- 0 elude but are not limited to immune complex diseases (such as systemic lupus erythematosus, arthritis, glomerulone- phritis, multiple sclerosis and AIDS) , Epstein Barr virus associated diseases (such as Sjogren's Syndrome, rheuma¬ toid arthritis, Burkitt's lymphoma, Hodgkinε disease, 5 virus (AIDS or EBV) associated B cell lymphoma, chronic fatigue syndrome, and parasitic disease states (such as viral infection following allograft transplantation or AIDS) . 5 2.6. CELL-MEDIATED IMMUNE RESPONSES A variety of immune responses independent of the complement system are known to be mediated by specifically reactive lymphocytes. These responses may give rise to autoimmune diseases, hypersensitivity, or simply allergic - j reactions. Some examples of these responses include delayed-type hypersensitivity, allograft rejection, graft versus host disease, drug allergies, or resistance to infection. Autoimmune disorders may include atrophic gastritis, thyroiditis, allergic encephalomyelitis, gas- 1 5 trie mucosa, thyrotoxicosis, autoimmune hemolytic anemia, and sympathetic ophthalmia (Eisen, H.N. , 1979, Immunology, Harper and Row, Hagerstown, Maryland, pp. 557-595) . The foregoing discussion of "Background of the Inven¬ tion" is not to be deemed an admission that any of the 0 foregoing is available as prior art against the instant invention. 3. SUMMARY OF THE INVENTION The present invention is directed to compositions 5 comprising combinations of organic and biological com¬ pounds that suppress immune responses, exhibit anti-in¬ flammatory activity, and/or selectively inhibit comple¬ ment. The compositions of the invention synergistically or additively inhibit the expression of complement-related 0 functions, or suppress immune activity and/or exhibit anti-inflammatory activity. The compositions of the present invention have therapeutic utility in the amelio¬ ration of disease and disorders mediated by complement and/or immune activity. The present invention is based on the surprising discovery that an organic compound that has the ability to inhibit complement activity, exhibit anti-inflammatory activity and/or suppress immune responses, and a biologi- cal compound with one or more of the same properties, can act synergistically or additively. The organic compounds in the compositions of the invention are those compounds that exhibit immunosuppres¬ sive and/or anti-inflammatory activity and/or that inhibit complement activity. Such organic compounds include but are not limited to K-76, K-76 COOH and derivatives and synthetic analogs thereof, antimetabolites, antimitotic drugs, cyclosporine, steroids, and nonsteroidal anti- inflammatory agents. Many such compounds are known in the art and can be used in the synergistic composition[s] provided by the present invention. In a particular em¬ bodiment, the organic compounds include but are not limit¬ ed to substituted dihydrobenzofurans, substituted and unsubstituted spirobenzofuran-2(3H)-cycloalkanes, and their open chain intermediates which exhibit such immuno¬ suppressive, complement inhibiting or anti-inflammatory activity. In specific embodiments, such organic compounds interrupt the proteolytic processing of C5 to bioactive components, blocking the release of C5a. The biological compounds of the invention contain the conserved SCR motif, and are able to inhibit complement activity. In particular, biological compounds of the invention include, but are not limited to, Cir, Cls, Factor B, C2, Factor H, C4-BP, DAF, MCP, CRl, CR2, C6, C7, interleukin-2 receptor a chain, b2-glycoprotein 1, and factor XIII. The biological compounds exhibit at least one of the functions associated with complement receptors and/or SCR containing proteins. In specific embodiments, the biological compounds are fragments, fusion proteins, chimeras, analogues or derivatives of soluble complement receptors CRl or CR2. In specific embodiments, the compositions of the invention may be used for the treatment of autoimmune 5 disease or the many diseases associated with the "inap¬ propriate" activation of the complement system or inflam¬ mation. In further embodiments of the invention, the mixture of organic and biological compounds is specific to the treatment of diseases or disorders associated with CRl * 0 and CR2. A further embodiment of this invention includes the combined therapy that can be obtained by treating patients with disorders that are routinely treated with thrombolyt- ic agents such as tissue plasminogen activator, streptoki- 5 nase or urokinase (e.g. myocardial infarction patients) with a combination of the synergistic composition[s] of this invention and the thrombolytic compounds. The present invention is also directed to pharma¬ ceutical compositions comprising combinations of such or- 0 ganic and biological compounds or the salts thereof. 3.1 DEFINITIONS As used herein, the following abbreviations and terms shall have the meanings indicated: 5 n-BuLi = n-butyllithium t-BuLi = tert-butyllithium t-BuSLi = lithium tert-butylthiolate C = complement; carbon CHO = Chinese hamster ovary 0 CRl = complement receptor type l (the C3b/C4b receptor CR2 = complement receptor type 2 CPM = counts per minute EBV - Epstein Barr Virus Et 2 0 = diethyl ether hexamethylphosphoric triamide immunoglobulin G isopropanol infrared the monocarboxylic acid derivative of K-76 the sodium salt of the monocarboxylic acid derivative of K-76 LAH lithium aluminum hydride LHR long homologous repeat MOM methoxymethyl group NK natural killer NMR nuclear magnetic resonance PBL peripheral blood lymphocyte(s) PCC pyridinium chlorochromate PHA phytohemagglutinin PMN polymorphonuclear cells RLi alkyllithium SCR short consensus repeat THF tetrahydrofuran TLC thin layer chromatography TMEDA N, ,N' ,N'-tetramethy1- ethylenediamine TMS tetramethy1si1ane TriMEDA N,N,N' trimethylethylenediamine sCRl soluble complement receptor type 1 The term "bioisosteric" group, as used in the present invention, describes an alternative chemical group whose electronic configuration is substantially analogous with the group to be replaced such that the polarity and charge of the whole molecule do not change. However, variations in the size, number of atoms or electron structure of the bioisosteric group (or "bioisostere") are permitted which variations may affect its function. Bioisosteres may be acidic (e.g., capable of releasing a proton and, subse- e quently, bearing a negative charge), basic (e.g., capable of being protonated and, subsequently, bearing a positive charge) or neutral (e.g., not normally capable of func¬ tioning as an acidic or basic group) . Unless otherwise stated or indicated, the term "al- kyl" as used herein refers to methyl, ethyl, and n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl groups. The term "alkanol" denotes a compound derived from coupling an alkyl group and hydroxyl radical. Simi¬ larly, the term "alkoxy" refers to methoxy, ethoxy, n-pro- pyloxy, isopropyloxy, n-, iso-, sec, and tert-butoxy groups. The term "lower" refers to the numerical range of 1 to 4 carbon atoms and includes linear or branched skele¬ tons. Unless otherwise stated or indicated, the term "halo- gen" as used herein includes fluorine, chlorine, bromine, and iodine. Unless otherwise stated or indicated, a given struc¬ ture, formula, or nomenclature for the substituted dihy- drobenzofuran analogs of this invention shall subsume all stereoisomers thereof. Unless otherwise stated or indicated, a reference made to a final compound of the invention which is a carboxylic acid, is also meant to include the salt form of such carboxylic acid such as alkali and alkaline-earth metal salts obtained therefrom. 4. BRIEF DESCRIPTION OF THE FIGURES FIGURE 1 demonstrates the inhibition of complement- mediated hemolysis by the disubstituted spirobenzofuran compounds 62, 66, and 68. Inhibition is shown as a func¬ tion of compound concentration . FIGURE 2 shows results from the SRBC hemolytic assay in which the concentration of 68 was varied in the ab- c sence of added sCRl (open squares) . Also shown is the inhibition of hemolysis as a function of compound concen¬ tration at a constant concentration of sCRl of 220 ng/ml (filled squares) . FIGURE 3 shows the inhibition of hemolysis as a func- tion of sCRl concentration in the presence or absence of added compound 68. The concentrations of sCRl required for 50% inhibition of hemolysis are 230 ng/ml and 100 ng/ml in the presence and absence of 68 (43 μM) , respec¬ tively. Thus the apparent potency of sCRl to inhibit complement hemolysis is nearly doubled in the presence of 43 μM 68. 5. DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to compositions comprising combinations of organic and biological com¬ pounds that suppress immune responses, exhibit anti-in¬ flammatory activity, and/or selectively inhibit comple¬ ment. The compositions of the invention synergistically or additively inhibit the expression of complement-related functions, suppress immune activity and/or exhibit anti- inflammatory activity. As used herein, the term "synergistic" refers to an effect of activity that is greater than additive of the individual affects or activities of each component. That is, the composition of the invention has greater inhibito¬ ry activity than the biological compound or the organic compound alone at the same concentration as they are present in the composition, and greater than the additive activity of both compounds. Preferably, the activity of a composition of the invention can be about 10% greater than the additive activity of the compounds. More preferably, the activity of the composition is about 25% greater, and even more preferably about 50% greater than the additive activity of each compound. In a specific embodiment, the in vitro inhibitory activity of the biological compound is about 30% and the inhibitory activity of the organic compound is about 3%, resulting in an additive activity of 33%, while the inhibitory activity of the composition comprising both compounds is about 50%. Thus, the compo- si ion has about 50% greater activity than the additive effects of the compounds: 100 X [(50 - 33) / 33] = 50% In another specific embodiment In vitro, the biological compound is about 2-fold more active with the organic compound present than in the absence of the organic com¬ pound. In yet another specific embodiment in vivo, the composition has at least 45% greater activity than the additive activity of each compound. As used herein, the term "additive" refers to the unexpected ability of the compounds to complement each other's activity to greater effect in vitro or in vivo than either compound alone could achieve. For example, if the biological compound achieves at best 40% inhibition and if the organic compound at best 30% inhibition of detrimental complement activity, e.g., in myocardial infarction, in a composition of the invention, the com¬ pounds together achieve greater than 40% inhibition. The greater than 40% inhibition provides more effective thera¬ peutic effect than could otherwise be expected. T e composition of the invention, which comprises a combination of organic and biologic compounds, can exhibit its inhibitory activity at an effective concentration, as described in the Examples, infra. However, the composi¬ tion can be prepared at a much higher concentration, for later dilution in vitro or in vivo to achieve an effective concentration. The organic compounds in the compositions of the invention are those compounds which exhibit immunosuppres- c sive, complement inhibiting, and/or anti-inflammatory activity. Such organic compounds include but are not limited to K-76, K-76 COOH and derivatives and synthetic analogs thereof, antimetabolites, antimitotic drugs, cyclosporine, steroids, and nonsteroidal anti-inflammatory agents. In a specific embodiment, the organic compound is cyclosporin. Many such compounds are known in the art and can be used in the synergistic composition[s] provided by the present invention. In a particular embodiment, the organic compounds include but are not limited to substi- 5 tuted dihydrobenzofurans, substituted and unsubstituted spirobenzofuran-2(3H)-cycloalkanes, and their open chain intermediates which exhibit such immunosuppressive, com¬ plement inhibiting or anti-inflammatory activity. In particular, such compounds are partial analogs of the fungal metabolite K-76. In a particular embodiment, the organic complement inhibitors of the invention inhibit C5 activation, that is, the proteolytic generation of bioactive complement fragments C5a and C5b from C5. The compositions compris- ing such compounds have value in the treatment of preven¬ tion of diseases or disorders associated with undesirable or inappropriate activation of the complement system. In specific embodiments, the compositions of the invention containing such compounds can be used in the treatment of inflammatory disorders. They may also be used for the treatment of cardiovascular disease. The present invention also relates to compositions in which the organic compounds possess immunosuppressive activity. In particular, such compounds inhibit immune responses. In specific embodiments, the compositions of the invention can inhibit the killing activity of mononu- clear cells, lymphocyte proliferation and/or activation. The compositions of the invention comprising immunosup¬ pressive compounds can be valuable in the treatment of various immune disorders. Furthermore, the organic compounds in the composi¬ tions of the invention may possess one or more of the K-76-like activities described supra in Section 2.4 and in the references cited therein. In a particular aspect, the organic compounds are non-protein compounds. Biological compounds of the compositions of the invention include those that contain the conserved SCR motif, that are able to inhibit complement activity, and that have at least one of the functions associated with complement receptors and/or SCR containing proteins. In specific embodiments, these biological compounds have at least one of the functions associated with comple¬ ment receptors and/or SCR containing proteins and may be fragments, fusion proteins, chimeras, analogues or deriva¬ tives of soluble complement receptors CRl or CR2. The present invention is also directed to pharma¬ ceutical compositions comprising mixtures of such organic and biological compounds or the salts thereof. The compounds of the present invention, and the intermediates and methods used in their preparation, and their pharmacological compositions are described in detail below. 5.1. COMPOUNDS WHICH INHIBIT COMPLEMENT AND/OR SUPPRESS IMMUNE ACTIVITY The present invention relates to compositions com¬ prising'organic compounds which synergistically or additively inhibit complement and/or suppress immune activity. Such organic compounds include but are not limited to K-76, K-76 COOH, derivatives and synthetic analogs thereof, immunosuppressives such as antimetabolit- es (e.g., azathioprine, methotrexate) , antimitotic drugs (e.g., cyclophosphamide) , cyclosporine (cyclosporine A), 5 macrolides such as FK506 (Nature, 341:758(1989)); and anti-inflammatory agents such as steroids (e.g., predni- sone, adrenal steroids such as glucocorticoids and cortic- osteroids) , and nonsteroids (e.g., indomethacin, salicyla- tes such as aspirins, ibuprofen, and naprosyn (arylacetic 10 acid group) ) . Many such compounds are known in the art and can be used in the practice of the invention. Exam¬ ples of some such compounds are described below. Organic compounds suitable as a component of the composition of the invention are those capable of exhibit- 15 ing an immunosuppressive activity, in particular, comple¬ ment inhibition or anti-inflammatory activity, and includ¬ ing but not limited to inhibition of interleukin biosyn¬ thesis or modulation of autoimmune disease. For example, compounds such as 3-substituted-2-oxindole-l-carboxamides 20 that have been described as inhibitors of interleukin-1 biosynthesis (U.S. Patent No. 4,861,794), may be used advantageously in the present invention. Other compounds include low molecular weight heparin fragments as inhibi¬ tors of complement activation (U.S. Patent No. 4,847,338), 252-phenylimidazole (2,l-B)benzothiazoles or salts thereof (U.S. Patent No. 4,464,384), benzanilides (U.S. Patent No. 4,072,753), hydroxamic acids of alicyclic a inoacids (U.S. Patent Nos. 3,997,594 and 3,703,543), compounds which bind to a heparin receptor such as those described in EPA 0375,976, homogeneous concanavalin-A dimer (U. S. Patent No. 4,889,842), or dibenzothiophene and thioxanthene compounds like those described in European Patent Applica¬ tion No. 342,433. Moreover, additional compounds which may be useful in 35 the present invention include cyclosporin and cyclodextr- in, as described, for example, in Australian Patent Application No. 8,817.386. In very particular cases, an immunosuppressive protein and its monoclonal antibody, such as that used for treating transplant rejection and c immune suppression due to burns, cancer or trauma (See, for example, U. S. Patent No. 4,925,920) may be useful in the instant composition. In other instances, the component may be a specific oligopeptide, capable of blocking immune complex binding to immunoglobulin Fc, such as that described in U. S. Patent No. 4,752,601. Likewise, certain low molecular weight quinoline derivatives, including cyclosporin, may be used (See, European Patent No. 231,151; Section 10, infra) . Other representative compounds include, but are not limited to, azanaphthalenes (U. S. Patent No. 4,945,095), 8- substituted-9-benzylguanine derivatives (U. S. Patent No. 4,874,862), 2,8,9-trisubstituted purine derivatives (U. S. Patent Nos. 4,918,219 and 4,921,859), 2- hydroxyphenylimidazole-(2,1-b)-benzothiazole compounds and esters thereof as described in U. S. Patent No. 4,464,384, trimethyl hydroxypyrimidines (See, for example, South African (SU) Patent No. 933,096), amidinovinylphenyl esters useful as protease inhibitors (U. S. Patent No. 4,490,388), and l-amidino-4-piperidine carboxylates and propionates (U. S. Patent No. 4,433,152). Additional examples of organic compounds suitable as one of the components of the present invention include sterol glucoside compounds, such as those described in Japanese Patent Application No. 57,112,400, N-acylpeptides (ϋ. S. Patent No. 4,436,726), 6-substituted-6H- dibenzopyran derivatives (U. S. Patent No. 4,463,001), secoprostaglandin compounds, such as those described in Japanese Patent Application No. 56,150,038, 5-aryl-2- thiouracil derivatives, such as those described in Japanese Patent Appli.cti.on No. 56,059,762, anthrani.li.c acid derivatives, such as those described in Japanese Patent Application No. 88,027,337, and 3-deazaadenosine and its derivatives (U. S. Patent No. 4,309,419). In addition, other compounds may also be employed. for example, 2-aryl-lH-perimidines from 1,8- diaminonaphthalenes and aryl halides or aryl aldehydes may be used (U. S. Patent Nos. 4,224,326 and 4,294,964); decahydronaphthalenespirobenzofuran derivatives (U. S. Patent No. 4,229,466), N-hydroxyphenyl-L-glutamine or its sales (U. S. Patent Nos. 4,180,588 and 4,265,766) , aminopurine nucleoside compounds, such as those useful for treating Lupus erythematosis, haemolytic anemia, nephrosis, ulcerative colitis and other like diseases (U.S. Patent No. 4,081,534), certain isothioureas (See, for example, U. S. Patent Nos. 4,205,071, 4,294,854, 4,378,365, 4,490,387 and 4,649,138) and phenylalanyltyrosyl oligopeptides, such as those described in Patent Publication DE 2,617,202. In a particular embodiment, organic compounds for use in the composition of the invention comprise substituted dihydrobenzofurans of the general formula 3 and substi¬ tuted spirobenzofuran-2 (3H) -cycloalkanes of the general formula 4. The groups represented by R and R--R 4 include hydrogen and linear or branched lower alkyl groups having l to 4 carbon atoms as defined previously in Section 3.1, supra. In addition, Rι-R 4 may each independently represent halogen, amino, amidic, hydroxyl, hydroxyalkyl, alkyloxy, nitro, formyl, acetal, carboxyl, trifluoroacetyl, N-substituted lower alkyl carboxamide, substituted vinyl of 2-10 carbon atoms, or alkylidene cgroup of 2-20 carbon atoms. Other groups which may be represented independently by R 3 and R 4 include hydrocarbons of 4 to 24 carbon atoms which may be of medium-length, long-chain, linear, branched, cyclic, saturated, unsaturated, unsubstituted, or heteroatom substituted. Moreover R 3 , R 4 , and the carbon atom to which they are attached may form a cyclic hydro¬ carbon group of 5-24 carbon atoms which may include a five-, six-, or seven-membered saturated of unsaturated ring comprised exclusively of carbon and hydrogen, or in combination with a hetero-atom. The ring may be unsub¬ stituted or may contain extra-cyclic hetero-atom or hydro¬ carbon substituents. This invention also relates to synthetic open chain intermediate compounds of the general formula 5 wherein R, R-, and R 2 are defined as above for formulae 3 and 4. In addition, R 5 represents hydrogen, lower alkyl groups, or suitable hydroxyl protecting groups such as methoxymethyl, tetrahydropyrany1, 2-methoxypropyl, 2-methoxyethoxymethyl, triarylmethyl, benzyl, methylthiomethyl, or tert-butyl- dimethylsilyl group. The * z group encompasses chemical groups represented by Ri- , as defined above for formulae 3 and 4 as well as substituent cyclohexenylmethyl (5a) , limonenyl (5b) , and carvone-derived diol acetonide (5c) groups. Other compounds may also be derived from intermediates 5 and 5a-c, which are, in turn, converted to products of general formulae 3 or 4 as discussed in the following sections. Table V lists representative compounds which comprise the general formula 4 of the present inven¬ tion; this list is not intended to be comprehensive. " Substituents on the spirocyclohexane ring may also be present; e.g., 10-isopropenyl. The substituted dihydrobenzofuran and spiro- benzofuran-2(3H)-cyclohexane compounds of the present invention of the general formulae 3, , the synthetic intermediates of the general formula 5, and the salts thereof exhibit complement inhibition, as manifested by inhibition of complement-mediated C5a production and/or inhibition of complement-mediated hemolysis. The complement-inhibitory properties of the compounds of the invention can be evaluated by modification of known tech¬ niques, e.g. - the assay described in Section 6.36.1, infra. Treatment of the compounds of the present invention with appropriate basic reagents provides pharmaceutically acceptable salts thereof. 5.2. SYNTHETIC PROCESSES Processes are provided which comprise chemical steps for the synthesis of many of the organic compounds of the invention. 5 Such synthetic processes are diagrammed in detail in Scheme 1. The synthetic scheme depicted in Scheme 1 provides a shorter and more flexible route than one based on the syntheses of K-76 (Corey and Das J. Am. Chem. Soc. 1982, 104, 5551; McMurray et al. J. Am.Chem. Soc. 1985, 10107, 2712). Retrosynthetic evaluation of all the final compounds produced according to Scheme 1 ultimately results in two fragments; an aliphatic and an aromatic portion. These two portions can be joined by using regioselective ortho-lithiation and subsequent alkylation. 15 Two different strategies can then be employed for the production of the final compound of the invention: initial cyclization followed by aromatic functionalization or vice versa. 0 5 0 5 Scheme 1 11 14* (4. RR,-H. R 2 -COOH) (a) "BuLi, THf-TMEDA; (b) Cul; (c) RβX; (ύ) "BuLi Hmnβ/TMEDA; (»)E * (R 2 ); (T)H\ "PrOH; (;) Am__.ry* twin; (h) feuSU. l**Α Scheme I (cont'd) 20c 23a ( 3 , R. R , - H. R 2 . COOK Rj - C_--. R, » l-mβthyicyeehβχβn-4-v ) ( a) ft Bub , TriF / TMEDA: (b) Co', -c) R»X: (d) n BuU. HaxanaTMEDA. (β)E * (R ∑ ); ( H * . f*rOH. te)Amb β 'V«r ββ r.; h)m-x * . r** > (i)- ---%KOH 5.2.1. PREPARATION OF COMPOUNDS OF GENERAL FORMULA 6-CARBOXYL-4-SUBSTITUTED SPIRO[BENZOFURAN -2(3H)-CYCLOHEXANES1 6-carboxyl spiro[benzofuran-2 (3H)-cyclohexane] mole¬ cules that are also substituted at the 4 position to form a series of ether substituted derivatives can be prepared as shown in Schemes 2 and 3 below and as more fully de¬ scribed in a specific Example in Section 6 infra. Scheme 2 *i* \ R-CΞ, Λ-lb B-CH.CE. Scheme 3 a) R.CsHsCH j - _,) R ■ π-βu - c j R« C^r - d ) R-HGCH j CH-- e ! R-o-NO-C«r-._ - _..R=- D-HO-C S~4 s i -to - m. These compounds and the salts thereof exhibit complement inhibition as manifested by inhibition of complement mediated hemolysis. Other complement-inhibitory properties of these compounds can be evaluated by numerous complement assay techniques that are known in the art. Substituents for R in position 4 of compounds of the general formula 4 include where R is a hydrogen atom or a lower alkyl group (e.g. CH 3 -, CH 3 CH--, n-Bu-) , a func- tionalized lower alkyl group (e.g. HOCH 2 CH 2 -) , a benzyl or substituted benzyl group, a phenyl or substituted phenyl group (e.g. C 6 H 5 CH 2 -, C 6 H 5 -, p-N0 2 C 6 H 4 -, p-CHOC 6 H 4 -, p-N0 2 C 6 H 4 -, and p-NH 2 C 6 H 4 -) . In addition the OR at position 4 of compounds of the general formula 4 can instead be H-. Preferred substitutions at position 4 are exemplified by compounds 44b, 55c and 55e, which are more effective in inhibiting complement mediated hemolysis than is K76C00H. 5.2.2. PREPARATION OF COMPOUNDS OF GENERAL FORMULA 6, 7 -DISUBSTITUTED -4-METHOXYSPIRO rBENZOFURAN-2 (3H) -CYCLOHEXANESi . . . 6, 7-dιsubstιtuted spιro[benzofuran-2 (3H) -cyclohexane] molecules can be prepared as shown in Schemes 4 and 5 below and as more fully described in Section 7 infra. Scheme 4 56 58 59 65 67 64 63 Sche e 5 11 T* .29^ eorr.. 63 SM i !9T<- (t l 1 - c 0 lib 65 6S In addition the compounds of this section can also be substituted at position 4 of the benzene ring to form 5 trisubstituted analogues. These di- and tri-substituted compounds and the salts thereof exhibit complement inhibition as manifested by inhibition of complement mediated hemolysis. Other complement-inhibitory properties of these compounds can be evaluated by numerous 0 complement assay techniques that are known in the art. The R j and R 2 groups can be any combination of the following: a hydrogen atom, a carboxyl group, a formyl group, a hydroxymethyl group, an N-(lower alkyl) carboxamide group, a trifluoroacetyl group, a carbalkoxy 5 group, a halide group, a substituted vinyl group of 2-10 carbon atoms, an alkylidene group of 2-20 carbons, a methyl ketone group, other lower alkyl ketone group, an aryl ketone group, a trifluoroacetyl group, a sulfonamide group, an imide group, a tetrazole group, a tertiary 0 aliphatic amine group, an oxazoline group, an amidine group, or a hydrazone group. Specific substitutions for RI and R2 groups include -CHO, -CH-OH, -COOH, COCF-, SO-NH- and tetrazole, oxazo¬ line, imide or CH 2 NMe- derivatives. Substitutions at posi- b tions 6 and 7 can be cyclic compounds as exemplified by co pounds 62 and 68. The presence of polar groups in the 6 and 7 positions appears to affect complement inhibition activity, perhaps because such polar groups interact with regions of the complement receptors, c Specific compounds include compounds of general formula 4 in which R can vary generally as described above; R- is a carboxyl group or a bioisosteric acid group (such as sulfonamide, imide, or tetrazole) or a bioisos¬ teric basic group (such as a tertiary aliphatic amine, oxazoline, amindine, or hydrazone) , or a bioisosteric neutral group (such as trifluoroacetyl) ; R 2 is a formyl group or a bioisosteric group such as methyl ketone (ace- tyl) , other alkyl ketone, aryl ketone or other similar group. A preferred organic compound of the compositions of this invention is the 4,6,7 trisubstituted spiro[benzofuran-2(3)H cyclohexane] compound 68, which exhibits a relatively large amount of complement inhibi¬ tion in the hemolysis assay. Even more inhibitory com- pounds can be produced by combining the optimal subsit- utions at the 4 position with the 6,7 disubstitutions present in 68. In an even more preferred embodiment the organic compound of the compositions is 6-carboxyl-7- formyl-4-phenoxyspirobenzofuran-2(3H)-cyclohexane] . 5.3. BIOLOGICAL COMPOUNDS WHICH INHIBIT COMPLEMENT Biological compounds within the scope of this inven¬ tion include complement receptors or soluble members of the complement receptor family that contain the conserved SCR motif that are able to inhibit complement activity. The SCR is one of the most characteristic structures of the complement system. Biological compounds containing SCRs include, but are not limited to, Cir, Cls, Factor B, C2, Factor H, C4-BP, DAF, MCP, CRl, CR2, C6, C7, inter- leukin-2 receptor α chain, 02-glycoprotein 1, and factor XIII. The production and use of derivatives, analogues, and peptides related to CRl and CR2 are also envisioned, and within the scope of the present invention. Such functions include but are not limited to binding of C3b and/or C4b, in free or in complex forms, promotion of phagocytosis complement regulation, immune stimulation, ability to act as a factor I cofactor, promoting the irreversible inactivation of complement components C3b or C4b (Fearon, D.T., 1979, Proc. Natl. Acad. Sci. U.S.A. 76:5867; Iida, K. and Nussenzweig, V., 1981, J. Exp. Med. 153:1138), and/or by the ability to inhibit the alternative or classical C3 or C5 convertases. Preferably a biological compound with at least one functional activity of CRl or CR2 is used. Soluble constructs carrying complement binding sites may be used in synergistic compositions for the treatment of a number of diseases and disorders related to comple¬ ment dependent cellular activation, where administration of said constructs will inhibit activation of complement and the complement-dependent activation of cells. For example, in a specific embodiment, a soluble CRl molecule can be used which retains a desired functional activity, as demonstrated, e.g., by the ability to inhibit classical complement-mediated hemolysis, classical C5a production, classical C3a production, or neutrophil oxidative burst in vitro. To this end, an expression vector can be con¬ structed to encode a CRl molecule which lacks the trans¬ membrane region (e.g. , by deletion carboxy-terminal to the arginine encoded by the most C-terminal SCR) , resulting in the production of a soluble CRl fragment. In one embodi¬ ment, such a fragment can retain the ability to bind C3b and/or C4b, in free or in complex forms. Preferred expression vectors for soluble CRl are described in International Patent Publications WO 89/09220 and WO 91/05047. Constructs containing CR2 will compete with cell- bound CR2 for EBV, reducing binding of EBV to cells and inhibiting EBV infection. Such constructs will compete for C3dg and thereby inhibit B-cell activation. This effect is particularly important in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythe¬ matosus. By binding to iC3b, the constructs can inhibit phagocytosis by neutrophils and macrophages. The con¬ structs can also serve to reduce inflammation. The CRl or CR2 molecules or related derivatives, analogues, and peptides of the invention can be produced by various methods known in the art. The manipulations which result in their production can occur at the gene or protein level by any of numerous strategies known in the art (Maniatis, T. , 1982, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York) . In the production of a gene encoding an ana- logue, or peptide, care should be taken to ensure that the modified gene remains within the same translational read¬ ing frame, uninterrupted by translational stop signals, in the gene region where the desired complement-specific activity is encoded. In a particular embodiment, fusion proteins, consisting of a molecule comprising a portion of the CRl or CR2 sequence plus a non-CRl sequence, can be produced. Additionally, the CRl or CR2 gene can be mutated in vitro or in vivo, to create and/or destroy translation, initiation, and/or termination sequences, to create muteins possessing single or few amino acid substitutions, to create alternatively glycosylated fragments by altering sites for glycosylation or to create variations in coding regions and/or form new restriction endonuclease sites or . . . . destroy preexisting ones, to facilitate further in vitro modification. Any technique for mutagenesis known in the art can be used, including but not limited to, .in vitro site-directed mutagenesis (Hutchinson, C. , et al., 1978, J. Biol. Chem. 253:6551), use of TAB linkers (Pharmacia), c etc. Manipulations of the CRl or CR2 sequence may also be made at the protein level to produce a derivative of CRl or CR2. Any of numerous chemical modifications may be carried out by known techniques, including but not limited to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease NaBH 4 ; acetylation, formylation, oxidation, reduction; metabolic synthesis in the presence of tunicamycin; etc. In addition, analogues and peptides related to CRl or 5 CR2 can be chemically synthesized. For example, a peptide corresponding to a portion of CRl which mediates the desired activity (e.g. , C3b and/or C4b binding, immune stimulation, complement regulation, etc.) can be synthe¬ sized by use of a peptide synthesizer. Specific modifications of the nucleotide sequence of CRl or CR2 can be made by recombinant DNA procedures that result in biological compounds encoding proteins having multiple LHR or SCR sequences or portions of LHR. Such valency modifications can alter the extent of specific complement related activities. Further modifications include the generation of chimeric molecules containing portions of the CRl or CR2 LHR or SCR sequences attached to other molecules whose purpose is to affect solubility, pharmacology or clearance of the resultant chimeras. Such chimeras can be produced either at the gene level as fusion proteins or at the protein level as chemically produced derivatives. Chimeric molecules comprising portions of immunoglobulin chains can contain Fab or (Fab') 2 molecules, produced by proteolytic cleavage or by the introduction of a stop codon after the hinge region in the heavy chain to delete the F c region, or different immunoglobulin isotypes. Depending upon the ultimate therapeutic use of the fusion protein, it may be preferable to maintain the F c region of a non-complement activating isotype in the immunoglobulin portion of the chimeric protein to provide F c receptor-mediated clearance of the complement activating complexes. Other molecules that may be used to form chimeras include, but are not limited to, proteins such as serum albumin, heparin, or immunoglobulin, polymers such as polyethylene glycol or polyoxyethylated polyols, or proteins modified to reduce antigenicity by, for example, derivatizing with polyethylene glycol. Suitable molecules are known in the art and are described, for example, in U.S. Patents 4,745,180, 4,766,106 and 4,847,325 and references cited therein. Additional molecules that may be used to form derivatives of the biological compounds or fragments thereof include protein A or protein G (International Patent Publication #WO87/05631 published September 24, 1987 and entitled "Method and means for producing a protein having the same IgG specificity as protein G"; Bjorck et al. , 1987, 24:1113-1122; Guss et al., 1986, EMBO J. 5:1567-1575, Nygren et al. , 1988, J. Molecular Recognition 1:69-74). 5.4. COMBINATIONS OF ORGANIC AND BIOLOGICAL COMPOUNDS WHICH INHIBIT COMPLEMENT The biological compounds in the compositions of this invention may possess multiple activities that may not be possessed by the organic compounds. Advantages of the organic compounds are that they are of low molecular weight and can be delivered by some systems that may not be readily applicable to the biological compounds, e.g., orally. Thus, delivery of the organic compounds to the patient may be accomplished by a different route than delivery of the biological compounds to the patient. Since there can be a synergistic effect observed with the combination of both organic and biological compounds, c it may be possible to administer lower doses of both com¬ pounds than would be required for either compound alone to achieve the same activity. Such lower doses would be important in situations where toxicity is a problem. Alternatively, the effect of the compounds of the composition can be additive to achieve a therapeutic result that neither drug alone, even at its most potent dose, could achieve. Mixtures of the Organic and Biological Compounds. Appropriate mixtures of the organic compounds and bio- logical compounds can be achieved in vitro or in vivo to produce inhibition of complement activity. The compounds can be administered simultaneously or sequentially. Phar¬ maceutical compositions comprising synergistically or additively effective combinations of the inhibitive organic and biological compounds or the salts thereof are provided by the present invention. Such compositions comprise a therapeutically effective amount of the composition comprising biological compound (or an analogue, derivative, or fragment thereof) , and the organic compound, and a pharmaceutically acceptable carrier. Such a carrier includes but is not limited to saline, buffered saline, dextrose, and water. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anaesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients will be supplied either separately or mixed together in unit dosage form or concentrated in a hermetically sealed container such as an ampoule or sachette i.ndi.cati.ng the quantity of active agent in activity units. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade 'Water for Injection 1 or saline. Where the composition is to be administered by injection, an ampoule of sterile water for injection or saline may be provided so that the ingredients may be mixed prior to administration. A pharmaceutical pack comprising one or more containers filled with one or more the ingredients of the pharmaceutical composition is also within the scope of the invention. Direct Chemical Modification of the Biological Compound. As an alternative to mixing the organic and biological compounds to provide compositions to treat patients with complement mediated disease, it is also possible to chemically modify the biological compound by linking the small molecular weight organic compound di¬ rectly to it. Such chemical linkage of a small organic compound to a polypeptide can be accomplished by known methods in the art involving either direct attachment to the polypeptide through a reactive group on the organic compound and/or the polypeptide (e.g. through lysine, tyrosine, or cysteine groups of the protein) or through the use of linker compounds that provide a short and flexible arm between the organic compound and the polypep¬ tide. Linkers can be used to give flexibility to the binding and action of the organic compounds and include both reversible and irreversible linkers. Reversible linkers can be used where it is desirable for the chemi- cally modified biological compound to act as a prodrug whose activity is modulated as the reversible linkers reverse and thereby release the attached organic compounds. The extent of attachment of organic compounds need to be determined to optimize either the activity of the prodrug or the amount of activity obtained from the synergism between the organic and biological compounds. An example of a modification that can be made to the substituted spiro[benzofuran]-cyclcoalkane compounds to c achieve a reactive group that can be used to attach to a biological compound is given below in Scheme 6 for perillyl alcohol derivatives of the BCD-ring spiro[benzo- furan]-2(3H)-cyclohexanes. Scheme 6 70 71 72 76 77 78 The perillyl alcohol groups are made to the B cyclohexane ring, since the 4, 6, and 7 positions of the D ring of the spirobenzofuran are important in complement activity. Once the series of perillyl alcohol derivatives have been c produced, the alcohol group can be further modified to create a reactive group capable of covalently attaching to specific amino acids on the biological compound. The exact modification of the alcohol group is dependent upon which amino acid or amino acid derivative of the biological protein is chosen as the site of attachment. Such modifications of alcohols and of amino acids are well known in the art. Since the structure of the SCR units of complement receptor proteins involve cysteines, lysine or tyrosine groups would be preferred over cysteines as attachment sites. Specific examples of perillyl alcohol derivatives have been tested and do exhibit complement inhibition activity. 5.5. ABILITY TO INHIBIT COMPLEMENT The synergetic mixtures of the invention can be assayed by any techniques known in the art in order to demonstrate their complement inhibiting activity. Such assays include but are not limited to the following in vitro tests for the ability to inhibit complement system activity or to selectively inhibit the generation of complement-derived peptides: (i) measurement of inhibition of complement-mediated lysis of red blood cells (hemolysis) (ii) measurement of ability to inhibit formation of C5a and C5a des Arg and/or measurement of abili¬ ty to inhibit formation of C3a and C3a des Arg. Those organic and biological compounds which are demonstrated to have significant complement-inhibiting activity can be therapeutically valuable for the treatment or prevention of diseases or disorders such as those described in Section 5.7, infra. Any organic compound as described in Section 5.1 that has the ability to inhibit any one of the activities associated with complement and any biological compound as described in Section 5.3 that 5 has any one of the activities associated with complement receptors, which in combination, behave synergistically or additively compared to the effects of either alone, is within the scope of this application. Activities normally associated with CRl and CR2 are well documented in the art .JO and include but are not limited to those activities and assays described in International Patent Application number PCT/US89/01358, published October 5, 1989 as W089/- 09220 and entitled "The Human C3b/C4b Receptor (CRl)"; Weissman et al. , 1990, Science 249:146-151; Fearon, D.T. 15 and Wong, W.W. , 1983, Ann. Rev. Immunol. 1:243; Fearon, D.T., 1979, Proc. Natl. Acad. Sci. U.S.A. 76:5867; Iida, K. and Nussenzweig, V., 1981, J. Exp. Med. 153:1138; Klickstein et al. , 1987, J. Exp. Med., 165:1095; Weiss et al. , 1988, J. Exp. Med., 167:1047-1066; Moore et al. , 201987, Proc. Natl. Acad. Sci. 84:9194; Moore et al., 1989, J.Biol.Chem. 264:20576) . For example, for CRl, such activities include the abilities in vitro to inhibit neutrophil oxidative burst, to inhibit complement-mediated hemolysis, to inhibit C3a and/or C5a production, to bind 5 C3b and/or C4b, to exhibit factor I cofactor activity, and to inhibit C3 and/or C5 convertase activity. In particular, the following assays can be used to demonstrate complement inhibition. 0 5.5.1. DEMONSTRATION OF INHIBITION OF C3a AND C5a PRODUCTION The ability to inhibit complement can be tested by assaying for specific inhibition of C3a and C5a produc¬ tion. For all experiments, a single human serum pool, to 5 be used as a source of complement, is aliquoted and stored frozen at -70°C. Human IgG is heat-aggregated, aliquoted, and stored frozen at -70°C. For each experiment, serum aliquots are equilibrated at 37°C with varying concentrations of the compounds tested. The classical c complement pathway is initiated by the addition of aggre¬ gated human IgG. Control samples containing no IgG should be included. After a fixed reaction time of 10 minutes (determined in an earlier time-course study to provide a convenient time interval during which the production of C5a or C3a is nearly complete, i.e., greater than 90%), the levels of the released complement peptides (C5a or C3a) are determined, for example, by radioimmunoassay using commercially available radioimmunoassay (RIA) kits (C5a RIA, UpJohn Cat. No. 3250-02; C3a RIA, UpJohn Cat. No. 3245-01; C5a RIA, Amersham Cat. No. RPA.520; C3a RIA, Amersham RPA.518) in modified procedures. Since a competitive immunoassay is used, complement peptide (C5a and C3a) concentrations vary inversely with the counts. The Counts Bound (CB) for a sample can be de- fined as the total counts (in counts per minute, cpm) measured in the pellet minus the counts measured in a non¬ specific binding (NSB) control. The NSB control is a sample containing only tracer peptide ( 125 I-labelled) and second precipitating antiserum; it contains no C5a- or C3a-specific antiserum. The fraction inhibition is equal to the Counts Bound (CB) for a "sample," less the CB in the "sample with no added compound," divided by the CB for the "no IgG control" less the CB in the "sample with no added compound." INHIBITION = f fCB sample) - (CB no compound.1 [ (CB no IgG) - (CB no compound) ] 5.5.2. DEMONSTRATION OF INHIBITION OF COMPLEMENT-MEDIATED HEMOLYSIS The ability to inhibit complement can also be tested by assaying for inhibition of complement-mediated red cell lysis (hemolysis) . The inhibition of hemolysis is deter- mined as a function of composition concentration, or by varying the concentration of one compound while keeping the concentration of the other compound constant. The compositions to be tested are prepared in 0.1 M Hepes buffer (0.15 N NaCl, pH 7.4), and 50 μl are added to each well of a V-bottom microtiter plate. Human serum, used as the complement source, is diluted 1 to 500 in Hepes buffer, and 50 μl are added to each well. Next, commercially available sheep erythrocytes with anti-sheep antibody (e.g., Diamedix Cat. No. 789-001) are used as received and added at 100 μl/well to initiate the complement pathway leading to hemolysis. The plate is incubated for 60 minutes at 37°C and then centrifuged at 500 x g for 10 minutes. The supernatants are removed and placed in a flat-bottom microtiter plate. The extent of hemolysis is measured as a function of the sample absorbance at 410 nm. The maximal absorbance (corresponding to maximal hemolysis) , - ^ , is obtained from the absorbance value of an erythrocyte sample containing only human serum, A s , less the absorbance of a sample containing only the red cells, Q. Thus, A,.,,-. = AS -AQ. The difference between the absorbance of an erythrocyte sample containing both human serum and inhibitive composition, and the absorbance of a cell sample containing inhibitive composition only, is defined as A.,--,.-,.. The inhibition, IH, is expressed as the fraction A- ^ - ^ /- , and IH 50 is defined as the concentration of inhibitive composition, or the individual components varied, required to produce a value of IH = 1/2. 5.6. IMMUNOSUPPRESSIVE ACTIVITY The organic compounds of the compositions of the present invention can inhibit immune activity. In partic¬ ular, the compounds of the invention inhibit cell-mediated c immune function. For example, the compounds can suppress natural killer activity, inhibit the proliferation of peripheral blood lymphocytes, and/or inhibit the activa¬ tion of T lymphocytes in PBL culture. Any procedure known in the art may be employed to demonstrate immunosuppressive activity. Such procedures include but are not limited to in vitro assays for inhibi¬ tion of natural killer lysis of target cells, inhibition of proliferation of peripheral blood lymphocytes or inhi¬ bition of cell surface interleukin-2 receptor expression. In particular, the following assays can be used to demonstrate immunosuppressive activities. 5.6.1. DEMONSTRATION OF INHIBITION OF NATURAL KILLER ACTIVITY The composition can be tested for its effect on the ability of peripheral blood mononuclear cells to lyse NK- sensitive target cells, K562. NK activity is determined using 51 Cr-labeled K562 erythromyeloid leukemia cells as targets, and normal peripheral blood mononuclear cells as effector cells, in a four hour cytotoxicity assay. Effec¬ tor cells are isolated from fresh blood by Ficoll-Hypaque gradient centrifugation, and 100 μl of a 5 x 10 5 cell/well suspension are added to each well of a V-bottom microtiter plate. The composition is diluted in RPMI 1640 medium containing 10% fetal calf serum and dispensed at 100 μl per well. Target cells (K562) are labeled for 30 minutes with 100 μCi of 51 Cr, washed thoroughly and dispensed in a 20 μl volume at 10 4 cells/well. These cell concentrations result in an effector-to-target cell ratio of 50:1. The microtiter plate is centrifuged, e.g., at 50 x g for 5 minutes and incubated in a humidified chamber with 5% C0 2 at 37°C. After 4 hours, 100 μl are removed from each well and the radioactivity is measured with a LKB 1275 gamma counter. The percent specific lysis was calculated as follows: % specific lysis = [(EXP - SR)/(TOTAL - B) ] X 100 where EXP (experimental value) is obtained using effector and target cells; SR (the spontaneous release) is obtained from target cells incubated with media alone; TOTAL re¬ lease is obtained by hypotonic lysis in water; and B represents instrumental background. Means can be calculated from quadruplicate wells. Viability of the effector cells incubated with the test compound can be determined by trypan blue exclusion. 5.6.2. DEMONSTRATION OF INHIBITION OF PROLIFERATION OF PERIPHERAL BLOOD LYMPHOCYTES The proliferation of human peripheral blood lympho¬ cytes (PBL) in response to phytohemagglutinin (PHA, Well¬ come) or anti-CD3 monoclonal antibody (OKT-3, Ortho) can be assessed by the incorporation of 3 H-thymidine. The composition is diluted in culture media to the desired concentrations, human PBL are added to a final concentration of 10 6 cells/ml, and then either PHA (Wellcome) or anti-CD3 (OKT3, Ortho) antibody (final concentration, 1 mg/ml) are added to initiate proliferation. The final volume per sample can be 100 μl. The cells are incubated for 72 hours after stimulation, pulsed with 1 μCi 3 H-thymidine per sample for 4 hours, harvested, and 3 H-thymidine incorporation in DNA is counted in a scintillation counter. Separate experiments, conducted on samples exposed only to varying amounts of the composition without the stimulant, can show that the PBL are viable, as determined visually by trypan blue exclusion, in the concentration ranges of the composition that are used. Release of cell surface interleukin-2 receptor (IL- 2R) or CD8 antigen from lymphocytes is a correlate of T cell activation (Rubin et al. J. Immunol. 1985, 135, 3172- 77; Rubin et al. Fed. proc. 1985 44, 946; Fujimoto, J. et al. J. EXP. Med. 1983, 159. 752-66; Tomkinson, B. et al. 2d Annual Conference on Clinical Immunol. Washington, D.C, October 30, 1987). The level of IL-2R or CD8 protein released into the supernatant of the PBL cultures can be assessed by removing aliquots therefrom just prior to pulsing with 3 H-thymidine. Commercially available enzyme immunoassay kits (CELLFREE™ IL-2R, Cat. No. CK1020, T Cell Sciences, Inc., Cambridge, MA, or CELLFREE™ T8/CD8, Cat. No. CK1040, T Cell Sciences) can be used to determine the levels of the two analytes. 5.6.3. DEMONSTRATION OF INHIBITION OF CELL SURFACE INTERLEUKIN-2 RECEPTOR EXPRESSION The interleukin-2 receptor (IL-2R) is not detectable on the surface of resting T cells. Upon activation by specific antigens or mitogens, T cell proliferation is mediated by an autocrine mechanism whereby activated cells secrete interleukin-2 and express cell surface IL-2R (Meuer, S.C. et al. Proc. Natl. Acad. Sci. U.S.A. 1984, 81, 1509; Tsudo, M. , et al. J. Exp. Med. 1984, 160, 612- 617; Waldmann, T.A. , et al. J. Exp. Med. 1984, 160, 1450- 1466) . Compositions can be tested for their ability to inhibit T cell activation, as indicated by inhibition of cell-surface IL-2R expression. PBL cultures (1.5 ml; 24- well plates) are stimulated with PHA (1 μg/ml) for 72 hours in the absence or presence of varying amounts of the composition. Subsequently, the cells can be stained using luorescein isothiocyanate (FITC)-labeled anti-IL-2R antibody (Act-T-Set IL-2R, Cat. No. AA2009, T Cell Sciences, Inc., Cambridge, MA) and analyzed by flow cytometry (Ortho System 30) . 5.6.4. LACK OF INHIBITION OF CHO CELL PROLIFERATION The specificity of a composition's inhibitory activity upon immune cells can be demonstrated by assaying for the ability to inhibit the proliferation of Chinese hamster ovary (CHO) cells. Non-confluent CHO cells are allowed to proliferate for 4 hours in a 96-well plate at a volume of 50 μl per well in the absence or presence of the composition. The cells are pulsed with 0.5 μCi of 3 H- thymidine per sample for 4 hours, harvested, and the amount of incorporated 3 H-thymidine is determined by scintillation counting. The CHO cells allowed to proliferate in the presence of the composition can incorpotrate roughly the same amount of 3 H-thymidine as cells incubated in the absence of the composition, thus showing whether a composition of the present invention that inhibits activation and/or proliferation of PBL cultures does not inhibit mammalian cell proliferation. 5.7. THERAPEUTIC USES OF THE COMPOUNDS OF THE INVENTION The compositions of the invention which exhibit synergistic or additive complement, immune and/or inflammatory activity inhibition have therapeutic value in the prevention or treatment of various immune or inflammatory diseases or disorders. The combination of compounds of the invention may be administered to a patient for treatment of an immune disorder involving undesirable or inappropriate complement activity. In particular, an effective dose of an inhibitive mixture of compounds of the invention may be therapeutically applied to ameliorate or to prevent a detrimental effect caused by the activity of a component of the complement system (e.g. , C5a) or an inappropriately reactive immune system. An effective dose of the composition can be determined by one of ordinary skill, and depends on the choice of biological compound and organic compound as well as the relative amounts of each taking into account routine safety studies, toxicity studies, dose ranges and method of delivery, e.g., bolus, continuous, or repeated. In a particular embodiment, the dose of the organic compound can range from about 0.01 to about 500 mg/kg/day, and the dose of the biological compound can range from about 0.01 to about 500 mg/kg/day. One or the other compound can be provided as a bolus dose. In a specific embodiment the organic compound is cyclosporin A given intramuscularly at about 10 mg/kg/day, and the biological compound is soluble CRl given as a single intravenous bolus of 15 mg/kg. The diseases or disorders which may be treated by the compounds of the invention include but are not limited to those listed in Table III and described in Section 2.6 supra. In particular, those disorders associated with extended zones of tissue destruction due to burn- or myocardial infarct-induced trauma, and adult respiratory distress syndrome (ARDS) also known as shock lung can be treated by administration of an effective amount of the compositions of the invention. Detrimental nonspecific activation of the complement system, or unfavorable activation by the alternative pathway, can also be prevented or treated by administering compositions of the invention. In specific embodiments, mixtures of such compounds can ameliorate the acute pathological changes induced by specific or non-specific proteolytic processing of C5. Mixtures of such compounds refers either to the combi.ned admi.ni.stration of organic and biological compounds or to the administration of biological compounds that have been chemically modified to also contain organic compounds. Mixtures of the compounds of the invention may also c be used to modulate biologic or immune functions directly or indirectly mediated by the complement system, which can include but are not limited to those functions listed in Tables I and II, supra, and the in vivo correlates of the in vitro functions therein. In particular embodiments, the mixtures of the inhib¬ itive compounds can be used to treat inflammation asso¬ ciated with, for example, kidney stones, systemic lupus erythematosis (SLE) , nephrotoxic glomeronephritis, or multiple sclerosis (See, e.g.. Experimental Allergic Encephalomyelitis. A Useful Model for Multiple Sclerosis, A Satellite Conference of the International Society of Neurochemists, July 16-19, 1983, University of Washington, Seattle, Washington; Miyazaki, W. et al. Microbiol. Immunol. 1980, 24, 1091; Konno, S. and Tsurutuji, S. Sr. J. Pharmacol. 1983, 80, 269). In yet another embodiment, the mixtures of the com¬ pounds of the invention can be administered for treatment of tissue damage due to myocardial ischemia and reperfusion, resulting from neutrophils attracted by and activated by the complement system. Mixtures of the compounds of the invention may also be administered for the prevention or treatment of diseas¬ es or disorders caused or accomplished by increased lym¬ phocyte or disorders caused or accompanied by increased lymphocyte or natural killer activity, including but not limited to atrophic gastritus, thyroiditis, allergic encephalomyelitits, gastric mucosa, thyrotoxicosis, auto¬ immune hemolytic anemia, pemphigus vulgaris, sympathetic opthalmia, delayed-type hypersensitivity, rejection of allografts, graft-host reaction, organ transplant rejec- tion, other autoimmune disorders, and drug allergies. They can also be used to alleviate the adverse effects of complement activation caused by therapeutic intervention such as tissue plasminogen activator therapy or cardiopul- monary bypass. Various delivery systems (e.g. , encapsulation in liposomes, microparticles, or microcapsules, conjugation to specific molecules) are known and can be used for therapeutic delivery of the compounds. Methods of admin- istration include but are not limited to oral, intrader- mal, transdermal, intravenous, subcutaneous, intramuscu¬ lar, intraperitoneal, and intranasal routes. Such administration can be done in either bolus or repeat doses or continuously by infusion for instance. The organic and biological compounds can be delivered concurrently or sequentially, by the same or different routes. For exam¬ ple, in one embodiment, the organic compound can be taken orally, while the biological compound is administered intravenously. A further embodiment of this invention includes the combined therapy that can be obtained by treating patients with disorders (e.g. myocardial infarction patients) that are routinely treated with thrombolytic agents such as tissue plasminogen activator, streptokinase or urokinase with a combination of the compositions of this invention and the routinely administered thrombolytic compounds or a fibrinolytically active fragment, derivative, or modified version thereof. The usefulness of such a combined thera¬ py derives from the observation that the complement system is activated in disorders such as myocardial infarction or bypass surgery. The efficacy of a combined treatment could be substantially better than the thrombolytic treat¬ ment alone due to the ability of the complement inhibitory compositions to modulate the inappropriate and damaging complement acti.vation. The admi.nistration of the throm- bolytic and composition could be simultaneous or sequential or in different dose forms including combi¬ nations of oral dose forms with injectables to name just a few. The invention can be better understood by referring to the following examples which are given for illustrative purposes only and are not meant to limit the invention. 6. EXAMPLES OF 6-CARBOXYL-4-METHOXYSPIRO rBENZOFURAN-2C3H)-CYCLOHEXANES1 10 6.1. 6-CARBOXYL-4-METHOXYSPIRO[BENZOFURAN- 2(3H)-CYCLOHEXANE] SODIUM SALT (44a) AND 6-CARBOXYLATE-4-ETHOXY-SPIRO [BENZOFURAN-2(3H)-CYCLOHEXANE] SODIUM SALT (44b) Preparation of 33a-b. Methyl 3,5-dihydroxybenzoate 15 (10 g, 59.5 mmol) in dry acetone (200 ml) was added to dry K 2 C0 3 (8.2 g, 59.4 mmol). Dimethysulfate (for 33a; 7.5 g, 59.5 mmol) was added to the mixture, which was refluxed for 24 h. The mixture was filtered. Ether (200 ml) was added to the solution and was extracted with H 2 0 (100 ml x 2) . The organic solution was dried over MgS0 4 ; after remov¬ al of solvent, a crude product (9.0 g) was purified by chromatography (1:19 EtOAc/CH 2 Cl 2 ) to afford 33a as a white solid, yield 3.9 g (36%). mp 95-97°C; TLC R f 0.58 (1:9 25 EtOAc/CH 2 Cl 2 ) . NMR (CDC1 3 ) S 7.26-7.13 (m,2H) , 6.63 (m,IH) , 5.90 (S,1H,0H), 3.90 (s,3H), 3.80 (s,3H). For 33b: The procedure of 33a was modified by using diethyl sulfate as alkylating agent. 33b: mp 98-100°C; yield, 33%; TLC R f 0.51 (1:9 EtOAc/CH 2 Cl 2 ) . NMR (CDC1 3 ) δ 7.16-7.14 (m, 2H) , 306.62 (m,lH), 5.70 (s,lH,0H), 4.07-4.00 (q,2H), 3.90 (S,1H), 1.41-1.38 (t,3H). Preparation of 34a-b. NaH (50% in mineral oil; 2.2g, 91.7 mmol) was washed with hexane (40 ml x 2) under N 2 . Dry DMF (50 ml) was added and the mixture was cooled to 0°C. A 5 solution of 33a (7.6 g, 41.7 mmol) in dry DMF (40 ml) was added slowly to the mixture, and then stirred for 1 h at 25°C. The mixture was recooled to 0°C, and a solution of MOMC1 (3.7 g, 45.8 mmol) in dry DMF (20 ml) was added dropwise. The mixture was stirred at 25°C for 3 h. Ether (200 ml) was added to the mixture and washed with H 2 0 (100 ml x 4) . The ether portion was dried (MgS0 4 ) and concen¬ trated to give a pale yellow liquid as 34a; yield 8.2 g (87%). TLC R f 0.44 (CH 2 C1 2 ) . IR (neat) 2950, 1740, 1600, 1460, 1430, 1320, 1240, 1150, 1060, 1020, 770 cm '1 . NMR (CDC1 3 ) Δ7.32-7.31 (m,lH), 7.24-7.23 (m,lH), 6.79 (m,lH), 5.19 (s,2H), 3.90 (s,2H), 3.83 (s,3H), 3.48 (s,3H). For 34b: Procedure for 34a was modified. Yield, 95%; TLC R f 0.42 (CH 2 C1 2 ) . IR (neat) 2980, 1725, 1600, 1450, 1300, 1240, 1150, 1060, 1030, 770 cm "1 . NMR (CDC1 3 ) δ 7.30-7.29 (m,lH), 7.23-7.22 (m, IH) , 6.79-6.78 (m,lH), 5.19 (s,2H) , 4.09-4.02 (q,2H), 3.90 (s,3H), 3.48 (s,3H), 1.44-1.39 (t,3H). Preparation of 35a-b. LiAlH 4 (1.7 g, 44.2 mmol) was suspended in dry THF (150 ml) under N 2 and cooled to 0°C. A solution of 34a (10 g, 44.2 mmol) in THF (20 ml) was added dropwise. The mixture was stirred for 3 h at 25°C. Excess LiAlH 4 was carefully decomposed by additional ice. The mixture was filtered through silica gel and rinsed with ether (150 ml) . The solution was washed with H 2 0 (50 ml x 2) , dried and concentrated to give a colorless liquid as 35a, yield 6.7g (76%). TLC R f 0.38 (1:4 EtOAc/CH 2 Cl 2 ) . IR (neat) 3400, 2920, 1600, 1460, 1290, 1150, 1050, 925, 840cm-'. NMR (CDC1 3 ) Δ6.67-6.50 (m,3H), 5.16 (s,2H), 4.65 (s,2H), s,3H), 3.50 (s,3H), 2.56 (s,H,0H). 35b was pre- pared by the same procedure that used for 35a: yield, 90-92% ; TLC R f 0 . 52 ( 1 : 4 Et 2 0/CH 2 C1 2 ) . IR (neat) 3400 , 2940 , 1600, 1460, 1390, 1280, 1210, 1155, 1140, 1030, 925, 850 cm" 1 . NMR (CDCI 3 ) δ 6.80-6.57 (m,3H), 5.27 (s,2H), 4.70 (S, 2H) , 4.20-3 . 97 (q, 2H) , 3 . 50 (s, 3H) , 2. 00 (s , H, OH) , 1.50-1.33 (t, 3H) . Preparation of 36a-b. To a solution of 35a (2.2 g, 11.1 mmol) and imidazole (1.51 g, 22.2 mmol) in CH 2 C1 2 (50 5 ml) , was added slowly TBDMSCl (2.0 g, 13.3 mmol) dissolved in CH 2 C1 2 (10 ml) . The mixture was stirred at 25°C for 3 h. The mixture was washed with H 2 0 (100 ml x 2) , dried with (MgS0 4 ) and concentrated to give a colorless liquid, 3.3 g (94%) as 36a. TLC R f 0.8 (4:6 hexane/CH 2 C1 2 ) . IR (neat) 102940, 1600, 1460, 1365, 1300, 1250, 1210, 1190, 1150, 1100, 1055, 1025, 925, 835, 765 cm "1 . NMR (CDC1 3 ) δ 6.62-6.61 (m,lH), 6.58-6.56 (m,lH), 6.49-6.47 (t,lH), 5.15 (s,2H), 4.69 (S,2H), 3.78 (S,3H) , 3.47 (s,3H), 0.95 (s,9H), 0.10 (s,6H). For 36b: It was prepared by the 15 procedure that used for 36a; yield, 90-95%. IR (neat) 2940, 1600, 1460, 1390, 1370, 1290, 1250, 1150, 1100, 1030, 940, 840, 775 cm "1 . NMR (CDC1 3 ) δ 6.61-6.59 (m,IH) , 6.56-6.54 (m,lH), 6.48-6.47 (t,lH), 5.15 (s,2H), 4.67 (s,2H), 4.04-3.97 (q,2H), 3.47 (s,3H), 1.43-1.37 (t,3H), 200.94 (S,9H), 0.10 (s,6H). Preparation of 38a-b. To a solution of 36a (8.2 g, 26.2 mmol) and TMEDA (4.6 g, 39.4 mmol) in dry THF (200 ml) under N 2 at 0°C, n-BuLi (12 ml, 28.8 mmol) was added slowly. The solution was stirred 30 min at 0°C and then 5 1.5 h at 25°C. The solution was recooled to -78°C, Cul (7.5 g, 39.4 mmol) was added in one portion under positive N 2 stream. The mixture was stirred for 1.5 h at -78°C to -40°C. The mixture was recooled to -78°C, bromide 37 (5.5 g . 31.5 mmol) in THF (20 ml) was added dropwise, and the 0 mixture was stirred 4 h from -78°C to 25°C. Ether (200 ml) was added to the solution and washed with 20% NH 4 0H solu¬ tion until the aqueous layer was no longer blue, dried and concentrated to give a brown liquid (10.4 g) . Purification by chromatography (1:19 EtOAc/hexane) afforded a colorless 5 liquid as product 38a (8.7 g, 82%). TLC R f 0.70 (1:9 EtOAc/hexane) . IR (neat) 2930, 1610, 1590, 1460, 1430, 1260, 1200, 1100, 1080, 1030, 840, 780 cm -1 . NMR (CDCl 3 ) δ 6.68 (S,1H), 6.61 (s,lH), 5.22-5.18 (m,lH) , 5.13 (s,2H) , 5 4.71 (s,2H), 3.79 (s,3H), 3.44 (S,3H), 3.26 (S,2H), 1.97-1.88 (m,4H), 1.62-1.46 (m,4H), 0.94 (s,9H), 0.10 (s,6H). For 38b: Preparation was essentially that used for 38a. Yield, 60-62%. TLC R f 0.66 (1:9 EtOAc/hexane) . IR (neat) 2940, 1610, 1590, 1440, 1390, 1370, 1260, 1200, 101150, 1100, 840, 770 cm "1 . NMR (CDC1 3 ) δ 6.67 (S,1H), 6.58 (S,1H), 5.25-5.22 (m,lH), 5.13 (s,2H), 4.69 (s,2H), 4.03- 3.96 (q,2H), 3.44 (s,3H), 3.28 (s,2H), 2.00-1.89 (m,4H), 1.76-1.47 (m,4H), 1.39-1.35 (t,3H), 0.94 (s,9H), 0.10 (S,6H) . 15 Preparation of 39a-b. To the solution of 38a (7.2 g, 14.7 mmol) in THF (100 ml), (Bu) 4 NF (1.0 M in THF, 21 ml, 21 mmol) was added slowly at 25°C. Solution was stirred at 25°C for 2 h. Ether (100 ml) was added to the solution, then washed with 5% HCl (20 ml) and H 2 0 (100 ml x 2) . The 0 organic layer was dried and concentrated to give a color¬ less liquid as 39a, yield 6.0 g (90%) . TLC R f 0.53 (2:3 EtOAc/hexane) . IR (neat) 3380, 3920, 1610, 1590, 1450, 1430, 1400, 1200, 1160, 1110, 1080, 1030, 960, 925, 830 cm "1 . NMR (CDC1 3 ) δ 6.70 (s,lH), 6.61 (s,lH), 5.21-5.18 5 (m,lH), 5.14 (S,2H) , 4.62 (s,2H), 3.79 (S,3H), 3.44 (S,3H), 3.26 (S,2H), 2.00-1.87 (m,4H), 1.62-1.48 (m,4H) . For 39b: Preparation was essentially that used for 39a. Yield, 95%. TLC R f 0.58 (2:3 EtOAc/hexane) . IR (neat) 3400, 2940, 1610, 1590, 1440, 1390, 1200, 1150, 1120, 0 1070, 1030, 925, 820 cm - 1 . NMR (CDC1 3 ) δ 6.69 (s,lH), 6.58 (S,1H) , 5.30-5.25 (m,lH), 5.14 (s,2H), 4.59 (s,2H), 4.03- 3.96 (q,2H), 3.44 (s,3H) , 3.28 (s,2H), 2.00-1.88 (m,4H), 1.60-1.46 (m,4H), 1.39-1.35 (t,3H). 5 Preparation of 40a-b. To a mixture of PCC (5.5 g, 25.6 mmol) in CH 2 C1 2 (100 ml) at 25°C was added 39a (4.6 g, 15.7 mmol) in CH 2 C1 2 (50 ml) slowly. The mixture was stirred at 25°C for 4 h more. The mixture was filtered through silica gel and rinsed with EtOAc (40 ml). The solution was washed with H0 (50 ml) , dried (MgS0 4 ) and concentrated to give a yellow liquid which then was puri¬ fied by chromatography (3:7 EtOAc/hexane) to afford a pale yellow liquid as 40a, yield 4.2 g (92%). TLC R f 0.66 (3:7 EtOAc/hexane) . IR (neat) 2920, 1690, 1580, 1450, 1430, 1380, 1300, 1200, 1150, 1110, 1070, 1020, 920, 840, 740, 720 cm "1 . NMR (CDC1 3 ) δ 9.89 (s,lH), 7.23 (S,1H), 7.10 (S,1H), 5.22 (S,2H), 5.20-5.18 (m, IH) , 3.86 (s,3H), 3.46 (s,3H), 3.33 (S,2H), 1.98-1.88 (m,4H), 1.61-1.48 (m,4H). For 40b: Procedure of preparation was essentially that used for 40a. Yield, 75-81%. TLC R f 0.56 (1:4 EtOAC/hexane) . IR (neat) 2920, 1700, 1585, 1440, 1380, 1310, 1200, 1160, 1110, 1070, 1030, 960, 920, 845, 740, 720 cm "1 . NMR (CDCI 3 ) δ 9.88 (s,lH), 7.22 (S,1H), 7.08 (s,lH), 5.25-5.22 (m,lH), 5.21 (S,2H), 4.12-4.05 (q,2H), 3.47 (S,3H), 3.36 (S,2H), 2.00-1.89 (m,4H), 1.60-1.38 (m,4H), 1.43-1.39 (t,3H). Preparation of 41a-b. To a solution of 40a (4.2 g, 14.5 mmol) in 2-propanol (30 ml) and THF (15 ml) at 0°C was added slowly aq. 4N HCl solution (40 ml, 160 mmol) in 2-propanol (10 ml) . The solution was then stirred at 25°C for 16 h or until disappearance of 40a (by TLC) . The solution was extracted with ether (50 ml x 3) . The ether solution was washed with H,0 (30 ml) , then dried and con- centrated. The crude product was purified by chromatogra¬ phy (3:7 EtOAc/hexane) to afford 41a as white solid, yield 3 g (84%). mp 153-155°C. TLC R f 0.60 (3:7 EtOAc/hexane) . NMR (CDCI 3 ) δ 9.90 (S,1H), 7.04 (m,2H), 5.78 (s,lH,0H), 5.68-5.65 (m,2H), 3.90 (s,3H), 3.48 (S,2H), 2.09-2.01 (m,4H), 1.96-1.90 (m,4H). For 41b: Procedure of prepara¬ tion was essentially that used for 41a. Yield, 76%; TLC R f 0.34 (1:4 EtOAc/hexane). NMR (CDC1 3 ) δ 9.86 (s,lH), 7.00 (m,2H), 5.80 (s,lH,OH), 5.70-5.66 (m,2H), 4.13-4.06 5 (q,2H), 3.47 (s,2H), 2.05-2.02 (m,2H), 1.92-1.88 (m, 2H) , 1.63-1.54 (m,4H), 1.46-1.41 (t,3H). Preparation of 42a-b. Amberlyst 15 (10 g) was added in one portion to the solution of 41a (3.3 g, 13.4 mmol) in CH 2 C1 2 (100 ml) . The mixture was stirred at 25°C for 6 10 h. The mixture was filtered and the solution was washed with H 2 0 (100 ml) , dried and concentrated to give a crude product (3 g) which then was purified by chromatography (1:4 EtOAc/hexane) to afford 42a as a white solid (2.5 g, 76%). TLC R f 0.57 (1:4 EtOAc/hexane). IR (KBr) 2930, 1690, 151600, 1430, 1390, 1350, 1325, 1220, 1120, 1030, 920, 830, 805, 745 cm -1 . NMR (CDC1 3 ) δ 9.85 (S,1H), 6.93 (s,lH), 6.89 (S,1H), 3.88 (s,3H), 2.94 (s,2H), 1.86-1.64 (m,6H), 1.55-1.45 (m,4H). 13 C NMR (CDC1 3 ) δ 191.77, 160.53, 156.90, 138.19, 121.25, 105.65, 102.81, 90.36, 55.58, 38.48, 2037.23, 25.05, 22.95. For 42b: Procedure of preparation was essentially that used for 42a. mp 105-106°C. Yield, 82%; TLC R f 0.55 (1:4 EtOAc/hexane). NMR (CDC1 3 ) δ 9.84 (s,lH), 6.91 (S,1H), 6.87 (S,1H), 4.15-4.08 (q,2H), 2.95 (s,2H), 1.86-1.68 (m,6H), 1.53-1.45 (m,4H), 1.45-1.40 (t,3H). 25 Preparation of 43a-b. To a mixture of aq. 2N NaOH (30 ml) containing Ag 2 0 (2.4 g, 10.2 mmol) at 50°C, a solu¬ tion of 42a (1 g, 4.1 mmol) in EtOH (1 ml) and THF (5 ml) was added slowly. The mixture was stirred for 6 h at 50°C. The mixture was filtered and the aq. solution was washed 30 with ether (50 ml) . The aq. solution then was cooled to 0°C, and acidified with cone. HCl solution. The white precipitate that resulted was extracted with ether, dried and concentrated to afford a white solid as 43a, yield 0 . 75g ( 71%) . TLC R f 0 . 62 ( 1 : 9 : 10 MeOH /CH 2 C1 2 / EtOAc ) . NMR 5 (CDCI 3 ) δ 7. 16 (S , 2H) , 3 . 88 (s , 3H) , 2 . 94 ( s , 2H) , 1. 87-1. 65 (m,6H), 1.55-1.47 (m,4H) . For 43b: Procedure for prepara¬ tion was essentially that used for 43a. mp 190-192°C. TLC R f 0.62 (1:9:10 MeOH/CH 2 Cl 2 /EtOAC) ; yield,69%. IR (KBr) 3300-2400, 1675, 1595, 1430, 1350, 1325, 1265, 1210, 1120, 1100, 1030, 955, 860, 770, 740 cm '1 . NMR (CDC1 3 ) δ 7.14 (s,2H), 4.15-4.08 (q,2H), 2.95 (s,2H), 1.86-1.65 (m,6H), 1.60-1.45 (m,4H) , 1.45-1.40 (t,3H). 13 C NMR (CDC1 3 ) δ 172.17, 160.08, 155.61, 130.19, 120.54, 105.39, 104.77, 1090.04, 63.80, 38.49, 37.24, 25.09, 22.99, 14.84. Anal. Calcd for C 16 H 20 0 4 : C, 69.54; H, 7.30. Found: C,69.40; H.7.35. Preparation of 44a-b. Sodium hydride (50% in mineral oil, 0.58 g, 24.2 mmol) was washed with hexane (30 ml) 15 under N 2 , then dry ether (50 ml) was added. A solution of 43a (3.2 g, 12.2 mmol) in ether (150 ml) was added to above mixture slowly at 25°C. The mixture was stirred at 25°C for 6 h and a white precipitate resulted. The mixture was extracted with H 2 0 (50 ml) . The aqueous solution was 20 collected and freeze dried to give a white solid (3.2 g, 92%). NMR (D 2 0) δ 7.10 (S,1H), 6.92 (S,1H), 3.85 (s,3H), 2.82 (S,2H), 1.70-1.54 (m,6H), 1.44-1.33 (m,4H). For 44b: Procedure for preparation was essentially that used for 44a. Yield, 75%. NMR (D 2 0) δ 7.09 (s,lH), 25 6.90 (S,1H), 4.19-4.12 (q,2H), 2.89 (S,2H), 1.77-1.60 (m,6H), 1.50-1.40 (m,4H), 1.40-1.36 (t,3H). 0 5 6.2. SODIUM SALTS OF 4-BENZYLOXY-6-CARBOXYLSPIRO [BENZOFURAN-2(3H) -CYCLOHEXANE] (55a) 4-N-BUTYLOXY-6-CARBOXYLSPIRO [BENZOFURAN-2 (3H) -CYCLOHEXANE] (55b) 6-CARBOXYL-4-PHENOXYSPIRO [BENZOFURAN-2(3H) -CYCLOHEXANE] (55c) 6-CARBOXYL-4-(2 '-HYDROXYETHYLOXYSPIRO [BENZOFURAN-2(3H) -CYCLOHEXANE] (55d) 6-CARBOXYL-4-P-NITROPHENOXYSPIRO [BENZOFURAN-2(3H) -CYCLOHEXANE] (55e) 6-CARBOXYLSPIRO[BENZOFURAN-2 (3H) - CYCLOHEXANE] (55g) ; AND 6-CARBOXYL-4-P-CARBOXYLPHENOXYSPIRO ΓBENZOFURAN-2 OH. -CYCLOHEXANE1 (551) 10 Preparation of 55a-e,g,i. Procedure for preparation of these compounds was essentially that used for 44. NMR spectra for 55a-g are listed as below. 55a: NMR (D 2 0) £ 7.21-6.88 (s,7H), 4.85 (s,2H), 2.56 (s,2H), 1.43-1.06 (m,10H). 55b: NMR (D 2 0) δ 7.10 (s,lH), 6.90 (s,lH), 4.13 (t,2H), 2.93 (s,2H), 1.79-1.67 (m,8H), 1.50-1.41 (m,6H), 0.93 (t,3H). 55c: NMR (D 2 0) δ 7.20-6.80 (m,7H), 2.61 (S,2H), 1.60-1.12 (m,10H). 55d: NMR (D 2 0) δ 7.07 (s,lH), 6.92 (S,1H), 4.18 (t,2H), 3.93 (t,2H), 2.96 (s,2H), 1.68 20 (S, b, 6H) , 1.44(s, b, 4H) . 55e: NMR (D 2 0) δ 8.10-8.07 (d,2H), 7.13 (S,1H), 7.11 (s,lH), 6.98-6.95 (d,2H), 2.68 (S,2H), 1.70-1.23 (m,10H). 55i: NMR (D 2 0) 7.90-7.87 (d,2H), 7.14 (S,1H), 7.12 (s, IH) , 7.00-6.97 (d,2H), 2.76 (s,2H), 1.70-1.27 (m,10H). 55g: NMR (D 2 0) δ 7.45-7.42 (d,lH), 257.23-7.20 (m,2H), 2.93 (s,2H), 1.65 (s, b, 6H) , 1.41 (s,b, 4H) . 13 C NMR (D 2 0) δ 177.63, 160.05, 139.25, 133.68, 127.83, 124.60, 112.09, 93.64, 42.45, 38.96, 27.15, 25.37. 7. EXAMPLES 30 7.1. 2-(1•-CYCLOHEXENYL)METHYL-3- METHOXYMETHOXYANISOLE (9 ) The 3-methoxymethoxyanisole, 7, was obtained in 96% yield by the following procedure. A mixture of finely powdered anhdryous potassium carbonate (K 2 C0 3 ; 2.0 equiv) 5 and 3-methoxyphenol (1.0 equiv) in dry acetonitrile was stirred at 0°C for 15 minutes under nitrogen. To ensure that the reaction pH remained above six, 100 ml of aceto¬ nitrile were used per gram of phenol substrate. A catalytic amount of 18-crown-6 (0.12 equiv) was added, and the mixture was stirred an additional 15 minutes at 0°C. Neat chloromethyl methyl ether (1.5 equiv) was then intro¬ duced slowly. The suspension was allowed to warm up to ambient temperature and stirred for 6 hours. After this time, the mixture was recooled to 0°C, and one-half the original quantities of K 2 C0 3 , 18-crown-6, and CH 3 OCH 2 Cl were added. After another 4 hours of stirring at room tempera¬ ture, the suspension was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in eithyl either (Et 2 0) , washed with 5% sodium hydroxide (3 x 50 ml), concentrated in vacuo, and dis¬ tilled under reduced pressure. Compound 7 was obtained as a clear colorless liquid. Bp 45°C (0.15 mm Hg) . l K NMR (90 MHz, deuteriochloroform) δ 7.16 (IH, m) , 6.61 (3H, ) , 5.16 (2H, s) , 3.79 (3H, s) , and 3.49 (3H, s) in ppm downfifled from TMS. 13 C NMR (CDC1 3 ) δ 160.5,m 158.2, 129.7, 108.2, 107.3, 102.5, 94.3, 55.9, and 55.1 in ppm downfield from TMS. n-BuLi (1.1 equiv) was added slowly to a THF solution of compound 7 (1.0 equiv) and TMEDA (1.1 equiv), at 0°C under a nitrogen atmosphere. The solution was stirred at room temperature for 2-5 hours and then cooled to -78°C. Cuprous iodide (1.2 equiv.) was added all at once. The light gray suspension was warmed up to -40°C, and after stirring for 1.5 hours, turned into a green-gray color. The copper reagent was cooled to -78°C and allowed to react with a THF solution of freshly-prepared allylic bromide 6a (1.3 equiv) . The reaction mixture was allowed to warm up to ambient temperature gradually and stirred for up to 72 hours. The mixture was quenched and washed . . . . with a saturated aqueous solution of sodium bicarbonate until the aqueous layer became colorless. The organic layer was dried by passage through a plug of potassium carbonate and concentrated in vacuo to give a dark orange oil. The crude product was distilled under reduced pres- 5 sure to provide compound 9a in 69% yield. Bp 105°C (0.15 mm Hg) . Anal. Caqlcd. for C- 6 H 22 0 3 : C, 73.25; H, 8.45. Found: C, 73.13; H, 8.50. Η NMR (90 MHz, CDCL 3 ) f 7.03 (IH, t, J = 8Hz) , 6.71 (IH, d, J = 8 Hz) , 6.54 (IH, d, J = 8 HZ), 5.23 (IH, broad s) , 5.13 (2H, s) , 3.77 (3H, s) , 103.43 (3H, s) 3,32 (3H, broad s) , 1.95 (4H, m) , and 1.57 (4H, m) in ppm downfield from TMS. 13 C NMR (75 MHz, CDC1 3 ) δ 158.5, 155.8, 136.3, 126.7, 120.0, 118.1, 107.0, 104.6, 94.3, 55.8 (2C's), 30.9, 28.8, 25.3, 23.1, and 22.6 ppm downfield from TMS. IR (neat) 2940, 2840, 1595, 1470, 151440, 1260, 1160, 1105, 1070, and 1025 cm "1 . 7.2. 3-(1'-CYCLOHEXENYL)METHYL-2-HYDROXY-4- METHOXYBENZOIC ACID C12a. Compound 9a was metalated at the 4-position by the „ following procedure. A hexane solution of 9a (1.0 equiv) and TMEDA (1.1 equiv) was treated with a hexane solution of n-BuLi (1.1 equiv) added gradually at 0°C under. an atmosphere of nitrogen. The solution was stirred at room temperature for 3 hours and then cooled to -78°C. Li-Sa The aryllithium reagent Li-9a prepared by the above route was then exposed to a stream of dried carbon dioxide 5 gas bubbled through the -78°C solution for 0.5 hour. Carrying out this reaction at 0°C cuts the resulting yield in half. The mixture was allowed to warm up to room temperature while maintaining a steady stream of gas. The mixture was poured into water and extracted a few times ith 5% aqueous sodium hydroxide, then acidified to pH 1 with concentrated hydrochloric acid, and then extracted into ether. The crude product was back-extracted into Et 2 0 and the combined organic layers dried over magnesium sulfate. The solvent was evaporated and the residue redissolved in a minimum of boiling ether. The warm solution was allowed to cool slightly, and then hexane was added to the point of cloudiness. The mixture was then allowed to stand in the freezer. An off-white solid with a melting point of 161-163°C was harvested (66% yield) . Anal. Calcd. for C 15 H- 8 0 4 : C, 68.69; H, 6.92. Found: C, 68.75; H, 6.95. l E NMR (90 MHz, acetone-d 5 7.81 (IH, d, J = 9 Hz) , 6.62 (IH, d, J = 9 Hz) , 5.26 (IH, broad s) , 4.26 (2H, broad s) , 3.90 (3H, s) , 3.29 (2H, broad s) , 2.01 (4H, m) , and 1.57 (4H, m) in ppm downfield from TMS. 13 C NMR (acetone-d^) δ 173.0, 164.3, 162.1 (2C's), 136.5, 130.6, 121.0, 115.9, 106.5 (2C's), 103.3, 56.3, 30.0, 29.4, 25.8, 23.8, 23.2 (2C's). IR (KBr) 1650, 1610, 1500, 1455, 1265, 1185, and 1090 cm "1 . 7.3. 3-(1'-CYCLOHEXENYL)METHYL-2-HYDROXY-4- METHOXYBENZALDEHYDE fl2b. A solution of aryllithium reagent Li-9a, prepared in situ by the procedure described in Section 7.2, supra, was cooled to -12°C and treated with neat N,N'- dimethylformamide (1.5 equiv) added all at once. The mixture was stirred at room temperature for 3 hours. The mixture was then poured into water, saturated with sodium chloride, and extracted with Et 2 0. The organic layers were combined, dried (MgS0 4 ) , and concentrated in vacuo; the residue was recystallized from ether-hexane (See Section 7.2). The product was purified by column chromatography (silica, ether/hexane eluent) . Mp 48-49°C. Anal. Calcd. for C- 5 H--0 3 : C, 73.15; H, 7.37. Found: C, 73.22; H, 7.40. *H NMR (CDC1 3 ) $ 11.42 (IH, broad s) , 9.64 (IH, s) , 7.33 5 (IH, d) , 6.51 (IH, d) , 5.23 (IH, broad s) , 3.83 (3H, s) , 3.26 (2H, broad s) , 1.96 (4H, m) , and 1.56 (4H, m) in ppm downfield from TMS. 13 C NMR (CDC1 3 ) <S 194.7, 164.6, 161.3, 135.5, 133.8, 120.6, 115.9, 115.6, 103.1, 55.9, 29.9, 28.8, 25.2, 23.0, and 22.4 in ppm downfield from TMS. IR 10 (KBr) 2930, 2840, 1625, 1495, 1255, 1100, 800, and 640 cm -1 . 7.4. 7-CARBOXY-4-METHOXYSPIRO[BENZOFURAN- 2 (3H)-CYCLOHEXANE! (Ila) 1 .5_ A benzene solution of compound 12a (Section 7.2) was treated with dry Amberlyst ion-exchange resin (approxi¬ mately 3-4 g per gram of substrate, dried at 110°C under high vacuum) added in one portion. The mixture was stirred for up to 24 hours at room temperature and then 2 - filtered. The resin beads were washed thoroughly with fresh benzene and methylene chloride. The filtrates were combined, washed with water, dried over magnesium sulfate, and concentrated in vacuo. The product Ila was purified in 85% yield by column chromatography. Mp 192-194°C. 25 Anal. Calcd. for C 15 H 18 0 4 : C, 68.69; H, 6.92. Found: C, 68.52; H, 6.99. J H NMR (CDC1 3 ) δ broad roll centered at about 12.0 (IH), 7.85 (IH, d, J = 9 Hz), 6.50 (IH, d, J = 9 Hz), 3.89 (3H, s) , 2.94 (2H, s) , and 1.69 (10H, broad m) in ppm downfield from TMS. 13 C NMR (CDC1 3 ) 164.5, 160.6, 30158.7, 132.7, 113.6, 106.2, 104.5, 94.2, 55.7, 38.0, 37.1 (2C's), 24.9, and 23.2 (2C's) in ppm downfield from TMS. Figure 1 shows the infrared (KBr) spectrum. IR (KBr) 1660, 1615, 1445, 1435, 1385, and 1100 cm "1 . Cyclization was also accomplished by heating the phenol precursor in 5 either a 50/50 4N hydrochloric acid/isopropanol solvent mixture or a boron trifluoride etherate solution in THF. This latter cyclization procedure is not recommended for any phenol precursor other than 12a, however. 7.5. 7-FORMYL-4-METHOXYSPIRO[BENZOFURAN- 2f3H)-CYCLOHEXANE! flib) The phenol precursor 12b was treated with Amberlyst resin as described in the previous section. The product lib was isolated and purified by column chromatography (83% yield). Mp 62-63°C. Anal. Calcd. for C 15 H--0 3 : C, , 73.15; H, 7.37. Found: C, 73.22; H, 7.40. l K NMR (CDCL 3 ) δ 10.16 (IH, s) , 7.66 (IH, d, J = 9 Hz) , 6.46 (IH, d, J = 9 Hz), 3.88 (3H, s) , 2.88 (2H, s) , 1.4-2.0 (10H, broad m) . Compound lib is easily transformed to lie by a metal hydride reduction step or other means well-known in the art. 7.6. SYNTHESIS OF 6-CARBOXY-7-FORMYL-4-METHOXY- SPIRO[BENZOFURAN-2(3H)-CYCLOHEXANE (68) AND 6,7-DICARB0XYL-4-METHOXYSPIRO fBENZOFURAN-2( ~ 3H)-CYCLOHEXANE. (66) Preparation of 65: A solution of n-butyllithium in hexane (2.20 ml, 4.95 mmol) was added to a solution of N, N, N' trimethylethylenediamme (0.65 ml, 5.09 mmol) in THF (6 ml) at -20°C (CC1 4 /C0 2 ) . After 30 min, lib (1170 mg, 4.76 mmol, prepared as described in section 6.8 supra) in THF (4 ml) was added dropwise, followed 30 min later by n-BuLi (6.35 ml, 14.28 mmol). The resulting system was kept at -20°C for 24 h, and DMF was added (02.20 ml, 28.56 mmol) . After 24 h reaction period, the reaction products were partitioned between ether (4 x 50 ml) and brine (50 ml), chromatography of the extracts gave 65 (1135 mg, 4.14 mmol, 87%) as a solid mp. 129-131°C (recrystallized from hexane-ether) ; IR (KBr): 3000-2840, 1670, 1600, 1470, 1425, 1390, 1320, 1280, 1260, 1210, 1130, 1030, 890, 850, 770, 700 and 620 cm "1 ; *H NMR: 1.40-1.93 (s, 10H) , 2.93 (s, 2H) , 3.94 (s, 3H) 7.04 (s, IH) , 10.35 (s, IH) and 10.70 (S, IH) ; 13 C NMR: 22.9, 24.9, 37.2, 37.7, 56.0, 93.0, 103.6, 113.7, 120.2, 138.7, 160.3, 164.7, 188.6 and 192.6; Elemental analysis: calcd. C=70.06, H=6.61; obsd. C=69.84, 5 H=6.65. Preparation of 68: A 4 N solution of potassium hydroxide (52.92 mmol) was added dropwise to a stirred solution of compound 65 (2900 mg, 1058 mmol) in THF (10 ml) and water (7 ml) containing dissolved silver nitrate 10 (3777 mg, 22.22 mmol, 1.05 eq.) at room temperature. The reaction system was protected from direct light. After stirring for 2 h at room temperature, the solids were filtered and extensively washed with distilled water. 2N H 2 S0 4 was added until pH 3, and the reaction products were 15 extracted with ether (3 x 150 ml) , washed with brine (2 x 25 ml) and dried (MgS0 4 ) . Chromatography of the reaction products allowed the recovery of the starting material (1823 mg, 63%), compound 68 (326 mg, 1.12 mmol, 11%, 29% corrected yield) and compound 66 (614 mg, 2.01 mmol, 19%, 2051%). Compound 68: mp: 148-150°C (rec. from hexane-ether) ; IR (KBr): 3440, 3000-2840, 1740, 1630, 1450, 1345, 1290, 1270, 1150, 1105, 1010, 910, 860, 770, 690, cm "1 ; *H NMR (DMSO d 6 ) : 1.30-1.52 (m, 4H) , 1.62-1.81 (m, 6H) , 2.91 (s, 2H) , 3.87 (s, 3H) , 6.55 (bs, IH) , 6.85 (s, IH) , and 7.95 25 (bs, IH) ; 13 C NMR (DMSO d 6 ) : 22.52, 24.47, 36.57, 37.56, 55.86, 91.90, 96.33, 98.34, 120.60, 121.48, 128.99, 154.0- 3, 158.28, 168.42; MS (e/m, %) : 290 (M+, 89), 289 (72), 272 (100), 271 (88), 244 (65), 215 (78), 192 (98), 191 (95), 190 (65), 165 (93), 164 (82), 79 (89). Elemental 0 analysis: calcd. C=66.20, H=6.25; obsd. C=66.19, H=6.26. Direct Preparation of 66: To a solution 65 (153 mg, 0.56 mmol) in ethanol (5 ml), was added a solution of silver nitrate (222 mg, 1.30 mmol) in distilled water (1 ml), followed by KOH (3 ml, 2.99 mmol). The system was 5 stirred overnight at room temperature, shielded from the light, then it was filtered and the residue was carefully washed with water. The combined aqueous phases were extracted with ether, the aqueous phase was then acidified 5 and extracted with ether (3 x 25 ml); the combined organ¬ ic phases were dried and chromatographed to afford 66 (156 mg, 0.51 mmol, 91%) as a white solid mp. 188-190°C (recrys¬ tallized from acetone); IR (KBr): 3500-2400, 3000-2850, 1700, 1610, 1410, 1330, 1290, 1130, 1040, 1000, 930, 855, 0 750 and 660 cm -1 : *H NMR: (acetone d 6 ) : 1.40-1.90 ( , 10H) , 2.93 (s, 2H) , 3.91 (s, 3H) and 6.95 (s, 2H) ; l3 C NMR: (acetone d 6 ) : 23.5, 25.6, 37.6, 38.7, 56.1, 91.7, 105.1, 111.6, 118.8, 133.0, 157.8, 158.8, 167.2 and 186.1; Ele¬ mental analysis: calcd. C=62.74, H=5.92; obsd. C=62.66, 15 H=6.01. 7.7. GENERAL METHODS The melting points were measured with a Thomas Hoover apparatus, and are uncorrected; the infrared spectra were 0 obtained with the aid of a Perkm Elmer 28IB spectropho- tometer, the ! H NMR spectra were recorded in deuterio- chloroform, unless another solvent is specified, either at 90 MHz, in a Varian EM 390 apparatus, or at 300 MHz in a Varian VXR 300 spectrometer. The 13 C NMR were recorded at 5 75.4 MHz in a Varian VXR 300 spectrometer. All the sig¬ nals are reported in ppm, downfield from tetramethyl- silane, used as internal reference. The multiplicities of the signals are abbreviated as: s=singlet, d=doublet, t=-triplet, q=quartet, m=multiplet, b=broad signal. The low resolution mass spectra were obtained from a Finnigan 3221-F200 spectrograph, at 70 eV. The elemental analyses were done by Atlantic Mirolab, Inc. (Norcross, GA) . The HRMS was obtained at Massachusetts Institute of Technolo- gy- All the solvents and chemicals used were of good quality. In several cases, some of them were purified and/or dried following preestablished procedures. All the reactions were carried out under a dry nitrogen 5 atmosphere. The column chromatographies were done using MN Silica gel 60, under atmospheric pressure, using dif¬ ferent solvent systems (hexane-ether, hexane-ethyl ace¬ tate) , in which increasing quantities of the second sol¬ vent were periodically added to the first solvent, or on a 10 chromatotron. 1% acetic acid was added during the chro¬ matographies of carboxylic acids. 8. COMPLEMENT INHIBITION BY Dl- AND TRI- SUBSTITUTED SPIRO[BENZOFURAN- 2 C3H) -CYCLOHEXANES1 15 The 4 substituted spirobenzofuran compounds of exam¬ ple 7 and the disubstituted spirobenzofuran compounds of example 8 were tested for their capacity to inhibit complement-mediated lysis of sheep red blood cells (SRBC) . The compounds to be tested were diluted in 0.1 M Hepes 20 buffer (0.15 N NaCl, pH 7.4), and 50 μl were added to each well of a V-bottom microtiter plate. Human serum, used as the complement source, was diluted 1 to 100 in Hepes buffer, and 50 μl were added to each well. Next, commercially available sheep erythrocytes with anti-sheep 25 antibody (Diamedix Cat. No. 789-001) were used as received and added at 100 μl/well to initiate the complement pathway leading to hemolysis. The plate was incubated for 60 minutes at 37°C and then centrifuged at 500 x g for 10 minutes. The supernatants were removed and 30 . placed in a flat-bottom microtiter plate. The extent of hemolysis was measured as a function of the sample absorbance at 410 nm. The maximal absorbance (corresponding to maximal hemolysis) , A_- ax , was obtained mm from the absorbance value of an erythrocyte sample containing only human serum, A s , less the absorbance of a sample containing only the red cells, AQ. Thus, A,- ^ = A s - Q . The difference between the absorbance of an erythrocyte sample containing both human serum and inhibitive compound, and the absorbance of a cell sample containing inhibitive compound only, was defined as A,,-.,,-.. The inhibition, IH, was expressed as the fraction (A, ^ - Aii m pi e )/- / arια _ l H 5o was defined as the concentration of inhibitive compound required to produce a value of IH = 1/2. The results of the hemolysis assays are shown in Figure l and Tables V and VI. Table V Inhibition of Hemolysis by 6,7-disubstituted -4-methoxy spirofbenzofuran-2(3H)-cyclohexanes] Compound Mean IH S0 (+ SD) n= mM * The concentration of compound (± standard deviation) required to produce a value for hemolysis inhibition of 0.5. Table VI Inhibition of Hemolysis by 6-carboxyl-4-substituted s iro benzofuran-2 (3H -c clohexanes ** The sodium salt of 4-p-aminophenoxy- carboxyspiro[benzofuran-2(3H)-cyclohexane] By comparing the inhibition of hemolysis by compounds 62, 66 and 68 as shown in figure 1 and Table V, it can be seen that the particular arrangement of substitutents at positions 6 and 7 of the benzofuran ring is important for anti-hemolytic activity. By comparing IH 50 values for the position 4-substituted series as shown in Table VI (for example, the values for 55c, 55e, and 55h) , improvements in anti-hemoly ic activity can also be obtained through the optimal choice of the R substituent at position 4. In addition, several of the compounds (see for example, compounds 55c, 55e, 55h and 68 in particular) are more effective in inhibiting complement-mediated hemolysis than is K76COOH. It is anticipated that optimal substituents at position 4, combined with an optimal pair of sub¬ stituents at position 6 and 7, will result in even more potent complement inhibitors. 9. SYNERGISTIC INHIBITION OF COMPLEMENT BY SOLUBLE COMPLEMENT RECEPTOR TYPE I (SCRl) AND 6-CARBOXY-7-FORMYL-4-METHOXY- SPIROΓBENZOFURAN-2f3H)-CYCLOHEXANE] (68) Soluble recombinant human complement receptor type 1 (sCRl) has been demonstrated to inhibit both classical and alternative complement activation in in vitro assays (International Patent Publications W089/09220 and WO91/05047, both entitled "The Human C3b/C4b Receptor (CRl)" and Weisman et al. , 1990, Science, 249, 146-151). In particular, sCRl has been shown to inhibit complement mediated sheep red blood cell (SRBC) lysis with 50% inhibition observed at concentrations ranging from 0.l to 1.0 nM depending on the specific assay conditions (Weisman et al. , ibid.) . Certain spirobenzofuran-2(3H)cyclohexanes have been shown to inhibit complement in in vitro assays (see Sec¬ tions 6, 7 and 8 supra) . Specifically, the compound 6- carboxyl-7-formyl-4-methoxyspiro[benzofuran-2(3H)cyclo- hexane], referred to herein as compound 68, has been shown to inhibit complement mediated sheep red blood cell (SRBC) hemolysis with 50% inhibition observed at a compound concentration of 164 ± 76 μM (see Section 8, Table V supra) . To assess the effect of combining both sCRl and compound 68 on the ability to inhibit complement, com¬ binations of these inhibitors were evaluated in the SRBC hemolytic assay. FIGURE 2 shows results from the SRBC hemolytic assay in which the concentration of 68 was varied in the absence of added sCRl (open squares) . Also shown is the inhibition of hemolysis as a function of compound concentration at a constant concentration of sCRl of 220 ng/ml (filled squares) . At this concentration in the absence of added 68, sCRl inhibits hemolysis by 30% (y-intercept value, filled squares) . As can be seen from the figure, at low concentrations of 68 (< 250 μM) , the inhibitory effects appear to be greater than a simple addition of independent activities, indicating a syner- gistic effect. For example, 11 μM 68 alone results in 3% inhibition, 220 ng/ml sCRl alone results in 30% inhibi¬ tion, but the combination of the two at these same concen¬ trations results in 54% inhibition. As can also be seen from the graph, at higher 68 concentrations (> 700 μM) , the combination with sCRl results in less than additive effects. The explanation for apparent synergistic effects at low concentrations of 68 and competitive effects at higher concentrations is being studied further. FIGURE 3 shows the inhibition of hemolysis as a function of sCRl concentration in the absence and presence of added compound 68. The concentrations of sCRl required for 50% inhibition of hemolysis are 230 ng/ml and 100 ng/ml in the absence and presence 68 (43 μM) , respec¬ tively. Thus the apparent potency of sCRl to inhibit complement hemolysis is nearly doubled in the presence of 43 μM 68. 10. EXAMPLE: THE EFFECTS OF CYCLOSPORINE AND SCRl ON SENSITIZED CARDIAC ALLOGRAFT REJECTION The present example describes the use of an established model of sensitized allografting, an ACI-to- Lewis rat cardiac transplant in which the Lewis rat recipient is sensitized by prior ACI skin grafting, to assess the effects of the combined administration of cyclosporine A and sCRl on hyperacute rejection in vivo. 10.1. MATERIALS AND METHODS Cardiac allograft transplants to sensitized recipients were performed as described previously (Pruitt & Bollinger, 1991, J. Surg. Res. 50:350-355). Briefly, Lewis rats received three successive ACI rat skin grafts which resulted in high serum titers of ACI-specific antibodies. These hypersensitized Lewis rats then underwent heterotopic ACI cardiac allografting. Cyclosporine A was given intramuscularly at 10 mg/kg/day beginning two (2) days prior to transplant and continued until the time of graft rejection. sCRl was given as a single intravenous bolus at 15 mg/kg immediately prior to reperfusion of the graft. Control animals received an intravenous bolus of phosphate buffered saline (PBS) of a volume equivalent to those in the sCRl-treated group. Cardiac allografts were evaluated visually for the first 30 minutes following reperfusion and then by abdominal palpation every 1 hour until rejection. Rejection was defined as total cessation of cardiac graft contraction and was confirmed by direct visualization and histologic examination. 10.2. RESULTS AND DISCUSSION Table VII includes the allograft survival times (hours) for individual animals and the means and standard errors of the mean (SEM) for the various groups. The sCRl group yielded increased graft survival times approximately 10-fold longer than the control group. This is consistent with a significant prolongation previously observed in the same model for a lower sCRl does of 3 mg/kg (Pruitt & Bollinger, 1991, supra) . The cyclosporine A group yielded variable results which suggested a possible prolongation of graft survival. An earlier study in the same model showed that cyclosporine A (10 mg/kg/day; intramuscularly beginning on the day of transplantation and continuing until rejection) produced a significantly longer graft survival than did controls while an identical except lower dose (5 mg/kg/day) did not (Knechtle et al., 1985 J. Heart. Transplant. 4:541-545). Unexplained variation in results within a single treatment group including cyclosporine have previously been reported in this model, for example, when combined with total lymphoid irradiation (Harland et al., 1989 Transplant. Proc. 21:1118-1119). As shown in Table VII, the combined administration of cyclosporine and sCRl yielded graft survival times which were clearly longer than sCRl treatment alone. The combination cyclosporine and sCRl graft survival times suggest an improvement over the "cyclosporine alone" group although the variation in the latter group is large. Thus the combination of sCRl and cyclosporine results in an improvement relative to the administration of either compound alone. TABLE VII ACI-to-sensitized Lewis rat cardiac transplant 11. DEPOSIT OF MICROORGANISMS E. coli strain DK1/P3 carrying plasmid piABCD (desig- nated pCRl-piABCD) , encoding the full-length CRl protein, was deposited with the Agricultural Research Culture Col¬ lection (NRRL) , Peoria, Illinois, on March 31, 1988 and was assigned accession number B-18355. Chinese hamster ovary cell line DUX Bll carrying plasmid pBSCRlc/pTCSgpt clone 35.6, encoding a soluble CRl molecule, was deposited with the American Type Culture Collection (ATCC) , Rockville, Maryland, on March 23, 1989 and was assigned accession number CRL 10052. The present invention is not to be limited in scope by the examples or by the microorganisms deposited since the compositions and the deposited embodiments are intended as single illustrations of aspects of the invention and any embodiments which are functionally equivalent are within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims. Various references are cited herein, the disclosures of which are incorporated by reference in their entire¬ ties. 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Claims: WHAT IS CLAIMED IS: 1. A composition comprising: (a) a biological compound that is able to inhibit at least one activity of complement; and (b) a non-protein organic compound that is able to exhibit an immunosuppressive, complement inhibiting or anti-inflammatory activity. 2. The composition of claim 1 in which the biological compound has an SCR or derivative thereof. 3. The composition of claim 1 which composition produces a complement inhibiting activity, which activity is manifested by: (i) inhibition of complement-mediated lysis of red blood cells; or (ii) inhibition of formation of C5a or C5a des Arg; or (iii) inhibition of formation of C3a or C3a des Arg. 4. The composition of claim l in which the biological compound is selected from the group consisting of CRl, CR2, CR3, CR4 Factor I, Factor H and DAF. 5. The composition of claim 1 in which the biological compound is CRl or a fragment or derivative thereof. 6. The composition of claim 1 in which the biological compound is CR2 or a fragment or derivative thereof. 7. The composition of claim 5 or 6 in which the biological compound is a soluble receptor substantially lacking a transmembrane region. 8. The composition of claim 1 further comprising a thrombolytic agent or fibrinolytically active fragment or derivative thereof. 9. The composition of claim 8 in which the throm- bolytic agent is selected from the group consisting of tissue plasminogen activator, streptokinase, and uroki- nase. 10. The composition of any one of claims 1-8 in which the non-protein compound is selected from the group consisting of anti-inflammatory compounds, antimetabolites, anti-mitotics, cyclosporin-like compounds and macrolides. 11. The composition of claim 1 in which the non- protein compound is a spirobenzofuran. 12. The composition of claim 11 in which the spirobenzofuran is selected from the group consisting of 6-carboxy1-4-phenoxyspiro[benzofuran-2(3H)-cyclohexane] , 6-carboxyl-4-p-nitrophenoxyspiro[benzofuran-2(3H)- cyclohexane] , 4-p-aminophenoxy-6-carboxyspiro[benzofuran- 2(3H)-cyclohexane] , 6-carboxyl-7-formyl-4- methoxyspiro[benzofuran-2(3H)cyclohexane] , 6-carboxyl-7- formyl-4-phenoxyspiro[benzofuran-2(3H)cyclohexane] , K76, and K76 COOH. 13. A molecule comprising a biological compound covalently linked to a non-protein compound, which biological compound is able to inhibit at least one functional activity of complement and which non-protein compound is able to exhibit an immunosuppressive or anti- inflammatory activity or able to inhibit at least one functional activity of complement. 5 14. The molecule of claim 13 in which the biological compound has an SCR or derivative thereof. 15. The molecule of claim 13 in which the covalent - lO linkage is cleaved after administration in vivo to a subject. 16. The molecule of claim 13 which produces a complement inhibiting activity, which activity is 15manifested by: (i) inhibition of complement-mediated lysis of red blood cells; (ii) inhibition of formation of C5a or C5a des Arg; or 0 (iϋ) inhibition of formation of C3a or C3a des Arg. 17. The molecule of claim 13 in which the protein is selected from the group consisting of CRl, CR2, CR3, CR4, 5 Factor I, Factor H, and DAF. 18. The molecule of claim 13 in which the protein is a soluble receptor substantially lacking a transmembrane region. 0 19. The molecule of claim 13 in which the protein is CRl or a fragment or derivative thereof. 20. The molecule of claim 13 in which the protein is CR2 or a fragment or derivative thereof. 21. The molecule of claim 19 in which the protein is a soluble CRl fragment substantially lacking a transmembrane region. 22. A composition comprising the molecule of claim 13 and a thrombolytic agent or fibrinolytically active fragment or derivative thereof. 23. The composition of claim 22 in which the thrombolytic agent is selected from the group consisting of tissue plasminogen activator, streptokinase, and uroki- nase. 24. The composition of claim 13 in which the non- protein compound is selected from the group consisting of anti-inflammatory compounds, antimetabolites, anti- mitotics, cyclosporin-like compounds and macrolides. 25. The composition of claim 13 in which the non- protein compound is a spirobenzofuran or derivative or analog thereof. 26. The composition of claim 25 in which the spirobenzofuran is selected from the group consisting of compounds 6-carboxyl-4-phenoxyspiro[benzofuran-2(3H)- cyclohexane] , 6-carboxyl-4-p-nitrophenoxyspiro[benzofuran- 2(3H)-cyclohexane] , 4-p-aminophenoxy- carboxyspiro[benzofuran-2(3H)-cyclohexane] , 6-carboxyl-7- formyl-4-methoxyspiro[benzofuran-2(3H)cyclohexane] , K76, and K76 COOH. 27. A composition comprising: (a) a soluble CRl fragment substantially lack¬ ing a transmembrane region which exhibits at least one functi.onal acti.vi.ty of CRl, which functional activity is selected from the group consisting of the ability in vi¬ tro to inhibit complement-mediated hemoly¬ sis, to inhibit C3a and/or C5a production, to bind C3b and/or C4b, to exhibit factor I cofactor activity, and to inhibit C3 and/or C5 convertase activity; and (b) 6-carboxy-7-formyl-4-methoxyspiro[benzofu- ran-2(3h)-cyclohexane] . 28. A pharmaceutical composition comprising an effective amount of the composition of claim 1 or 26 and a pharmaceutically acceptable carrier, for use in the treatment of an immune disorder or a disorder involving undesirable or inappropriate complement activity. 29. A pharmaceutical composition comprising an effective amount of the molecule of claim 14 and a pharmaceutically acceptable carrier for use in the treatment of an immune disorder or a disorder involving undesirable or inappropriate complement activity. 30. The composition of claim 1 in which the non- protein compound is a compound of the general formula, 4: in which R represents a hydrogen atom, a lower alkyl group, a substituted lower alkyl group, a benzyl group, a substituted benzyl group, a phenyl group or a substituted phenyl group; R- and R 2 represent independently a hydrogen atom, a carboxylic acid group, a formyl group, a hydroxymethyl group, a N-(lower alkyl)carbamoyl group, a trifluoroacetyl group, a halide group, a vinyl group, a substituted vinyl group having up to 10 carbon atoms, an alkylidene group having up to 20 carbon atoms, an aliphatic acyl group, a substituted aliphatic acyl group, an aromatic acyl group, a substituted aromatic acyl group, a sulfamoyl group, an aminomethyl group, a N-(lower alkyl)aminomethyl group, a N.N-di(lower alkyl)aminomethyl group, a heterocyclic ring, an N-(acyl)carbamoyl group, an amidino group or a hydrazide group; R, and R 2 together with the carbon atoms to which they are attached may also represent a cyclic anhydride or lactone; or a pharmaceutically acceptable acid or base addition salt or ester thereof, provided that R, and R 2 do not both represent hydrogen atoms.

Inventors: KUNG PATRICK C, MARSH HENRY C JR, KUNG, PATRICK, C., MARSH, HENRY, C., JR.
Applicants: T CELL SCIENCES INC, T CELL SCIENCES, INC.
Application year: 1991
Main classification: C12N9
IPC codes: A61K31, A61K38, A61K45, A61P29, A61P37, C07K14