Abstract:
The invention relates to the use of peptides individually or in combination, for treating and/or preventing angiogenesis. It also relates to the use of peptide analogs or a combination of peptides referred to as MuJ-7 as anticancer drugs in restricting the tumor growth and spread by inhibiting tumor angiogenesis. MuJ-7, in addition inhibits metastasis through its antiangiogenic activity in all cancers. The invention also relates to a pharmaceutical composition containing either individual peptides or in combination, and methods of treatment of human beings and animals for curing and/or preventing angiogenesis.

Description:
This application is a continuation-in-part of application No. 08/727,679 filed on Oct. 8, 1996 now U.S. Pat. No. 6,156,725 and claims benefit of Ser. No. 60/080,433 filed Apr. 2, 1998. 
    
    
     FIELD OF INVENTION 
     The invention relates to the use of peptides individually or in combination, for treating and/or preventing angiogenesis. It also relates to the use of a combination of peptides referred to as MuJ-7 as an anticancer drug in restricting tumor growth and its spread by inhibiting tumor angiogenesis. MuJ-7, in addition inhibits metastasis through its antiangiogenic activity in all cancers. The invention also relates to a pharmaceutical composition containing either individual peptides or combinations of peptides, and methods of treatment of human beings and animals for curing and/or preventing angiogenesis. 
     BACKGROUND OF THE INVENTION 
     Angiogenesis is the growth of new microvessels. This process depends mainly on locomotion, proliferation, and tube formation by capillary endothelial cells. During angiogenesis, endothelial cells emerge from their quiescent state and can proliferate as rapidly as bone marrow cells, but unlike the bone marrow, angiogenesis is usually focal and of brief duration. Pathologic angiogenesis, while still a focal process, persists for months or years. The angiogenesis that occurs in diseases of ocular neovascularisation, arthritis, skin diseases, and tumors rarely terminates spontaneously and has until recently, been difficult to suppress therapeutically. Therefore, the fundamental goal of all antiangiogenic therapy is to return foci of proliferating microvessels to their normal resting state, and to prevent their regrowth 1 . 
     Although the molecular mechanisms responsible for transition of a cell to angiogenic phenotype are not known, the sequence of events leading to the formation of new vessels has been well documented 2,3 . The vascular growth entails either endothelial sprouting 3,4  or intussusception 5 . In the first pathway, the following sequence of events may occur: (a) dissolution of the basement of the vessel, usually a postcapillary venule, and the interstitial matrix; (b) migration of endothelial cells toward the stimulus; (c) proliferation of endothelial cells trailing behind the leading endothelial cell(s); (d) formation of lumen (canalization) in the cndothelial array/sprout; (e) formation of branches and loops by confluence/anastomoses of sprouts to permit blood flow; (f) investment of the vessel with pericytes; and (g) formation of basement membrane around the immature vessel 2,3 . New vessels can also be formed via the second pathway: insertion of interstitial tissue columns into the lumen of preexisting vessels. The subsequent growth of these columns and their stabilization result in partitioning of the vessel lumen and remodelling of the local vascular network 5,6 . 
     The rationale for antiangiogenic therapy is that progressive tumor growth is angiogenesis-dependent 8 . The switch to the angiogenic phenotype appears to be an independent event that occurs during the multistage progression to neoplasia 9 . The angiogenic switch itself, while relatively sudden and well localized, is nonetheless a complex process. This phenotype is currently understood in terms of a shift in the net balance of stimulators and inhibitors of angiogenesis, during which inhibitors are down regulated 10,11 . 
     Once new capillary loops converge toward a small in situ carcinoma or a microscopic metastasis, the tumor cells are bathed in additional survival factors and growth factors, not only from the circulating blood (perfusion effect) but also from vascular endothelial cells themselves (paracrine effect). The positive regulators of angiogenesis include at least 14 angiogenic proteins that have been discovered during the past 12 years and which have been sequenced and cloned 12 . Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) are the most well studied and are found in a majority of different types of human tumors. During the angiogenic switch one or more of these angiogenic stimulators is upregulated and it appears, however, that this up regulation of angiogenic stimulators is accompanied by down regulation of local tissue inhibitors of angiogenesis. 
     The paracrine stimulation of tumor cells by products from endothelial cells also operates in the other direction. Endothelial cell survival and growth are driven by tumor derived mitogens and motogens. These findings have led to a model of tumor growth in which the endothelial cell compartment and the tumor cell compartment interact with each other. They not only stimulate each other&#39;s growth, but if the endothelial cells are made unresponsive to angiogenic stimuli from the tumor cells, by administration of a specific endothelial inhibitor, both primary tumors 11  and metastatic tumors 10  can be held dormant, at a microscopic size. One could take advantage of this difference between endothelial cells and tumor cells by administering an angiogenesis inhibitor together with conventional cytotoxic chemotherapy up to the point at which the cytotoxic therapy would normally be discontinued because of toxicity or drug resistance. The angiogenesis inhibitor(s) could then be continued (for years), to maintain either stable disease or tumor dormancy 1 . Such combinations of antiangiogenic and cytotoxic therapy in tumor-bearing animals have been curative, whereas either agent alone is merely as inhibitor 15 . 
     Peptides/Proteins have previously been studied for antiangiogenic activity. Thrombospondin-1 (TSP-1) which is a naturally occurring inhibitor of angiogenesis, makes endothelial cells unresponsive to a wide variety of inducers. Both native TSP-1 and small antiangiogenic peptides derived from it show that this inhibition is mediated by CD36 16  (Dawson et al., 1997). Both IgG antibodies against CD36 and glutathione-S-transferase-CD36 fusion proteins that contain the TSP-1 binding site blocked the ability of intact TSP-1 and its active peptides to inhibit the migration of cultured microvascular endothelial cells. 
     The family of tissue inhibitors of metalloproteinases (TIMPs) are known to be specific inhibitors of matrix metalloproteinases (MMPs). The local balance between MMPs and TIMPs is believed to play a major role in extracellular matrix (ECM) remodelling during diseases such as cancer. TIMP-3 which is unique in being a component of ECM, inhibits endothelial cell migration and tube formation in response to angiogenic factors 17  (Anand-Apte et al., 1996). 
     The conditioned medium of human promyelocytic leukemia (HL6O) cells has been shown to contain a cell growth inhibitory factor, human cytostatin. Human cytostatin can inhibit endothelial cell proliferation, migration and microvessel tube formation on Matrigel-coated surfaces 18  (Yeung AK et al., 1996). Furthermore the anti-angiogenic effect of human cytostatin has been demonstrated on the chick chorioallantoic membrane. Human cytostatin can inhibit new blood vessel development, but cannot regress existing blood vessels. 
     Angiostatin, which is a 38 kD internal fragment of plasminogen is an antiangiogenic endothelial cell inhibitor and suppresses the growth of primary Lewis lung carcinoma in vivo 19  (Wu, Z. et al., 1997). 
     Thalidomide has recently been shown to antagonize basic fibroblast growth factor-induced angiogenesis in the rat corneal micropocket assay. It has been suggested that thalidomide elevates tumor hypoxia in the Lewis lung tumor, presumably via an antiangiogenic mechanism 20  (Minchinton AI et al., 1996). 
     One study examined the in vitro antiangiogeneic effects of the somatostatin analog octreotide on the growth of human umbelical vein endothelial cells (HUVEC) and vascular cells from explants of rat aorta cultured on fibronectin-coated dishes or included in fibrin gel. A total 10 −9 M octreotide reduced the mean uptake of  3 H-thymidine by HUVEC cells by 37% compared with controls. The 10 −8M  concentration of octreotide inhibited the proliferation of endothelial and smooth muscle cells growing on fibronectin by 32.6% and reduced the sprouting of cells from the adventitia of aortic rings in fibrin by 33.2% compared with controls, as measured by tetrazolium bioreduction and image analysis, respectively. These results demonstrate that octreotide is an effective inhibitor of vascular cell proliferation in vitro 21  (Danesi R. Del Taccam, Metabolism August 1996. 45(8 Supp 1: 49-50). In another experiment, somatostatin analogs SMS 201-995 and RC-160 were found to inhibit angiogenesis in the chick chorioallantoic membrane of the developing chicken embryo. Somatostatin analogues were associated in a dose-related fashion with both a greater percentage of inhibition of blood vessel growth and an increased grade of inhibition. It was hypothesized that inhibition of angiogenesis may be a mechanism responsible for the tumor regression observed in vivo following SMS or RC-160 therapy 22  (Woltering E A et al., J. Surg. Res. March 1991, 50(3): 245-251). 
     The proinflammatory neuropeptide, substance P, stimulated angio-genesis in an in vitro model using HUVECs cultured on a basement membrane (Matrigel) substrate. Substance P stimulated endothelial cell differentiation into capillary-like structures in a dose-dependent manner. Stimulation of endothelial cell differentiation is a newly recognized biological function of substance P 23  (Wiedermann, C. J. et al. Eur. J. Pharmacol. Mar. 18, 1996, 298 (3): 335-338). 
     The effects of Nitric oxide (NO) generation and endogenous production of NO elicited by substance P (SP) in angiogenic process were evaluated in the rabbit cornea (in vivo) and by measuring growth and migration of endothelial cells (in vitro). The NO synthase inhibitors given systemically inhibited angiogenesis elicited by [sar9]-SP-Sulfone. Capillary endothelial cell proliferation and migration produced by SP were abolished by pretreatment with the NO synthase inhibitors. Exposure of the cells to SP activated the calcium-dependent NO synthase. These data indicate that NO production induced by vasoactive agents, such as SP, functions as an autocrine regulator of the microvascular events necessary for neovascularization and mediates angiogenesis 24  (Ziche M. et al., J. Clin. Invest. November 1994, 94(5):2036-2044). 
     The angiogenic activity of four vasoactive peptides with a range of vasodilator and vasoconstrictor properties were investigated in a rat sponge model. 2 daily doses of vasodilator peptide VIP(10 pmol) when given with interleukin-1 alpha caused intense neovascularization which was inhibited by simultaneous administration of VIP(10-28), a specific VIP receptor antagonist. These data show that VIP possesses angiogenic activity and the blockade of VIP induced angiogenesis at the receptor level could provide a strategy for the management of angiogenic disease 25 . 
     Vascular endothelial cells are important in a variety of physiological and pathophysiological processes. The growth and functions of vascular endothelial cells are regulated both by soluble mitogenic and differentiation factors and by interactions with the extracellular matrix; however, relatively little is known about the role of the matrix. The neuropeptide bombesin, the bioactive lipid lysophosphatidic acid (LPA), and the cytokine tumor necrosis factor alpha, which signal through diverse mechanisms, were all able to activate MAPK to a much greater degree in fibronectin adherent cells than in suspended cells. Together, these data suggest a cooperation between integrins and soluble mitogens in efficient propagation of signals to downstream kinases. This cooperation may contribute to anchorage dependence of mitogenic cell cycle progression (Short S M et al, Molecular Biology of the Cell 9(8):1969-1980, 1998). 
     Cell adhesion to the extracellular matrix (ECM) has been implicated in apoptosis in anchorage-dependent cell types. It was recently found that a peptide derived from fibronectin (termed III14-2) inhibits the integrin-mediated cell adhesion to ECM. Using this antiadhesive peptide and a variety of ECM proteins, a critical role of the integrin-ECM protein interaction in apoptotic regulation of human umbilical vein endothelial cells (HUVEC) has been demonstrated. HUVEC in suspension undergoes apoptosis under the serum-free conditions, as judged by nuclear and DNA fragmentations (Fukai F et al, Experimental Cell Research, 242(l):92-99, 1998). 
     It has been previously shown that vasoactive intestinal polypeptide (VIP) induces endothelium-dependent relaxation of the human uterine artery. Non-competitive antagonism with methylene blue revealed that the pKa value for VIP-receptor complex was 8.10+/−0.10 (n=6) and the receptor reserve expressed as K-A/EC50 was 0.89+/−0.11, where pKa=log (10)K(A), and K-A is the dissociation constant of VIP-receptor complex (Jovanovic A. et al. Molecular Human Reproduction, 4(1):71-76, 1998). 
     Somatostatin (SRIF) exerts antiproliferative effects, and has recently been evaluated in clinical trials for the prophylaxis of restenosis following coronary angioplasty. 3 SRIF (0.1-1000 nM) caused a concentration-dependent inhibition of the bFGF-stimulated regrowth in CHO-K1 cells expressing human sst(2) (h sst(2)) or sst(5) (h sst(5)) receptors (pIC(50)=8.05+/−0.03 and 8.56+/−0.12, respectively). SRIF (0.1-1000 nM) was able to inhibit the bFGF-stimulated re-growth (pIC(50)=7.98+/−24 and 8.50+/−0.12, respectively). (Alderton F et al, British Journal of Pharmacology, 124(2): 323-330, 1998). 
     Substance P (SP) was analyzed in rat brain endothelium cultures after cytokine stimulation. SP secretion was found after stimulation with high doses of interleukin-l beta (IL-1 beta) and tumor necrosis factor alpah (TNF-alpha). Under cytokine stimulation, part of SP was bound to brain endothelial cell surface, suggesting the existence of an autocrine network for this neuropeptide. SP regulates cellular processes in the CNS, placenta and vasculature include permeability, inflammation, mitogenesis and transformation. Increased SPR mRNA level in response to E-2 were linearly related to increased [H-3]SP binding to the SPR (Villablanca A C et al, Molecular &amp; Cellular Endocrinology, 135(2):109-117, 1997). 
     SUMMARY OF INVENTION 
     The present invention provides pharmaceutical compositions for treating cancer angiogenesis and cancer metastasis. The invention provides a method of treating angiogenesis, cancer and cancer metastasis employing a pharmaceutically effective dosage of a combination of peptides or individual peptides. It is an object of this invention that a combination of peptides used is known as MuJ-7. Individual constituent peptides of MuJ-7 and pharmaceutically acceptable additives can be used. The present invention provides a pharmaceutical composition useful for killing or inhibiting multiplication of tumor cells as well as cancer cells. The pharmaceutical composition may also be useful in preventing, inhibiting, or modulating the hypersecretion of VIP, somatostain, bombesin, Substance P, or a combination of VIP, somatostatin, bombesin, or Substance P. 
     The composition may suitably comprise, consist of, or consist essentially of a therapeutically effective combination of peptide analogs of somatostatin, VIP, bombesin, and Substance P. The peptide analogs are described in more detail below, but constituents functionally interchangeable with those specifically described may also be employed in the claimed pharmaceutical composition. More particularly, the pharmaceutical composition may suitably comprise, consist of, or consist essentially of an analog of somatostatin and at least four peptides selected from the group consisting of a first analog of VIP, a second analog of VIP, a third analog of VIP, analog of somatostatin another analog of somatostatin, an analog of bombesin, and an analog of Substance P. More particularly, the composition may suitably comprise, consist of, or consist essentially of a therapeutically effective combination of peptide SOM 2  (an analog of somatostatin) and at least four of the following peptides: VIP 1  (a VIP anatagonist), VIP 2  (a VIP) receptor binding inhibitor), VIP 3  (a VIP receptor antagonist), SOM 1  (a somatostatin analog (also abbreviated “CTOP.” which is derived from the first letters of the following four amino acids: Cys 2 , Tyr, Orn 5 , and Pen 5 ), BOM 1  (a bombesin antagonist), and SP 1  (a Substance P antagonist). In a preferred embodiment, a pharmaceutically acceptable carrier, diluent, or solvent is used. The invention provides a method of treatment for humans, mammals, or other animals suffering from cancer or other tumors. The invention also provides a method of treatment for humans, mammals, or other animals suffering from hypersecretion of VIP, somatostatin, bombesin, Substance P, or a combination of VIP, somatostatin, bombesin, or Substance P. The method may suitably comprise, consist of, or consist essentially of administering a therapeutically effective dose of the pharmaceutical composition so as to prevent, inhibit, or modulate the hypersecretion of VIP, somatostatin, bombesin, Substance P, or a combination of VIP, somatostatin, bombesin, or Substance P. 
     In addition, the compositions may comprise, consist essentially of or consist of one or more of peptides identified below as DT-11; DT-12; DT-13; DT-14; DT-15; DT-16; DT-18; DT-19; DT-23; DT-24; DT-26; DT-27; DT-31; DT-33; DT-34; DT-62A; DT-62B; DT-71 which are peptide analogs. 
     DETAILED DESCRIPTION 
     We have observed that VIP (vasoactive intestinal peptide), somatostatin, substance P, and bombesin are secreted by at least some human tumor and cancer cells and that there are binding sites for these peptides on these cells. Specifically, out of a number of peptide growth regulators studied by indirect immunofluorescence, the four peptides (i.e., vasoactive intestinal peptide (VIP), somatostatin, Substance P, and bombesin) were shown to bind, to tumor cells. (Herein, the terms “peptide growth regulators”, and “peptides” each refer to VIP, somatostatin, Substance P, and bombesin). It may be that there is an autocrine mechanism for cell proliferation where the peptides are secreted by tumor cells and transduce a signal through specific receptors on the same cell type leading to cell proliferation. As will be described in more detail below, the effects of the analogs of somatostatin, VIP, bombesin, Substance P on the tumor cell growth and survival were studied using different assay systems. The amino-acid sequences of the seven analogs (VIP 1 , VIP 2 , VIP 3 , SOM 1 , SOM 1 , BOM 2  and SP 1 ) are disclosed in U.S. application Ser. No. 08/727,679. 
     
       
         
               
               
               
               
             
           
               
                   
               
               
                 Code 
                 Name 
                 Sequence 
                 SEQ ID NO: 
               
               
                   
               
             
             
               
                 VIP 1   
                 VIP antagonist 
                 (Lys-Pro-Arg-Arg-Pro-Tyr-Thr-Asp-Asn-Tyr-Thr- 
                 SEQ ID NO: 1 
               
               
                   
                   
                 Arg-Leu-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr- 
               
               
                   
                   
                 Leu-Asn-Ser-Ile-Leu-Asn-NH 2 ) 
               
               
                 VIP 2   
                 VIP receptor binding 
                 (Leu-Met-Tyr-Pro-Thr-Tyr-Leu-Lys) 
                 SEQ ID NO: 2 
               
               
                   
                 inhibitor 
               
               
                 VIP 3   
                 VIP receptor antagonist 
                 (His-Ser-Asp-Ala-Val-4-Cl-D-Phe-Thr-Asp-Asn-Tyr- 
                 SEQ ID NO: 3 
               
               
                   
                   
                 Thr-Arg-Leu-Arg-Lys-Gln-Leu-Ala-Val-Lys-Lys- 
               
               
                   
                   
                 Tyr-Leu-Asn-Ser-Ile-Leu-Asn-NH 2 ) 
               
               
                 SOM 1   
                 Somatostatin analog 
                 D-Phe-Cys-Tyr-D-Trp-Om-Thr-Pen-Thr-NH 2   
                 SEQ ID NO: 4 
               
               
                   
                 (CTOP) 
               
               
                 SOM 2   
                 Somatostatin analog 
                 Ala-Gly-Cys-Lys-Asn-Phe-Phe-D-Trp-Lys-Thr-Phe- 
                 SEQ ID NO: 5 
               
               
                   
                   
                 Thr-Ser-D-Cys (disulfide bridges: 3-14) 
               
               
                 BOM 1   
                 Bombesin antagonist 
                 D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHEt 
                 SEQ ID NO: 6 
               
               
                 SP 1   
                 Substance P antagonist 
                 D-Arg-Pro-Lys-Pro-D-Phe-Gln-D-Trp-Phe-D-Trp 
                 SEQ ID NO: 7 
               
               
                   
                   
                 Leu-Leu-NH 2   
               
               
                   
               
             
          
         
       
     
     As will be explained in more detail below, the combination of these seven analogs is known as MuJ-7. The analogs were synthesized manually and using a conventional peptide synthesizer. An example of a combination within the scope of the invention comprises SOM 2 , VIP 1 , VIP 2 , VIP 3 , SOM 1 , BOM 1 , and SP 1 . A combination hereinafter referred to as MuJ-7, was prepared using the following seven peptide analogs: (1) VIP 1 , (the VIP antagonist) having a molecular weight of approximately 3464.9 and a concentration of approximately 10 −7  M; (2) VIP 2 . (the receptor binding inhibitor) having a molecular weight of approximately 1027.55 and a concentration of approximately 10 −8 M; (3) VIP 3 , (the VIP receptor antagonist) having a molecular weight of approximately 3342.09 and a concentration of approximately 10 −8 M; (4) SOM 1  (the somatostatin analog (CTOP) having a molecular weight of approximately 1061.59 and a concentration of approximately 10 −9 M; (5) SOM 2 , (the analog of somatostatin) having a molecular weight of approximately 1637.0 and a concentration of approximately 10 −8 M; (6) BOM 1  (the bombesin antagonist) having a molecular weight of approximately 983.55 and a concentration of approximately 10 −8 M; and (7) SP 1  (the Substance P antagonist) having a molecular weight of approximately 1515.83 and a concentration of approximately 10 −8 M. The preceding sentence sets forth the preferred concentrations of the seven analogs comprising MuJ-expected that MuJ-7 would be effective if the concentration of each of the seven analogs ranged from approximately 10 −6 M to approximately 10 −12 M. 
     MuJ-7 may be prepared in the following way. A stock solution of each of the seven peptide analogs is prepared with a pH of approximately 7.0 to approximately 7.4. Although sterile phosphate buffered saline was used to prepare the stock solutions for the testing described below, other diluents may be used such as RPMI 1649, buffered saline, isotonic NaCl, Ringer&#39;s solution, water (for injection) distilled water, polyethylene glycol (near or in water), 2% Tween in water, dimethylsulfoxide to 50% in water, propylene glycol (neat or in water), balanced salt solution, glycerol, and other conventional fluids that are suitable for intravenous administration to obtain a pH in the range of approximately 7.0 to approximately 7.4 for each stock solution, the pH can be adjusted by using 1 N HCl for lowering the pH or 1 N NaOH for raising the pH, although the conventional agents for adjusting the pH can be used, the concentration of the peptide analog in each stock solution is approximately 10 −3 M. 
     Aliquots of the seven peptides analogs are mixed together such that the MuJ-7 formulation contained approximately equal weights of each of the seven peptide analogs. In MuJ-7, approximately, the concentration of VIP 1 , is 10 −7 M; the concentration of VIP 2  is 10 −8 M; the concentration of VIP 3  is 10 −8 M; the concentration of SOM1 is 10 −9 M; the concentration of SOM 2  is 10 −8 M; the concentration of BOM1 is 10 −8 M; and the concentration of SP 1  is 10 −8 M. In one exemplary embodiment, the pH of the MuJ-7 solution may range from about 7.0 to 7.4. To obtain a pH in this range, the pH can be adjusted by using 1 N NCl for lowering the pH or 1 N NaOH for raising the pH, although other conventional agents for adjusting the pH can be used. 
     In addition, a number of novel peptides also exhibited an antiangiogenic effect. These peptides are: 
     
       
         
               
               
               
             
           
               
                   
               
             
             
               
                 DT-11 
                 Aib-Met-Tyr-Pro-Thr-Tyr-Aib-Lys-OH 
                 (SEQ ID NO: 8) 
               
               
                 DT-12 
                 D-Leu-Met-Tyr-Pro-Thr-Tyr-Aib-Lys-OH 
                 (SEQ ID NO: 9) 
               
               
                 DT-13 
                 Leu-Met-Tyr-Pro-Thr-D-Tyr-Leu-Lys-OH 
                 (SEQ ID NO: 10) 
               
               
                 DT-14 
                 Leu-Met-Tyr-Pro-Thr-Tyr-D-Leu-Lys-OH 
                 (SEQ ID NO: 11) 
               
               
                 DT-15 
                 Leu-Met-D-Tyr-Pro-Thr-Tyr-D-Leu-Lys-OH 
                 (SEQ ID NO: 12) 
               
               
                 DT-16 
                 D-Leu-Met-Tyr-Pro-Thr-Tyr-D-Leu-Lys-OH 
                 (SEQ ID NO: 13) 
               
               
                 DT-18 
                 Aib-Met-Tyr-Pro-Thr-Tyr-Deg-Lys-OH 
                 (SEQ ID NO: 14) 
               
               
                 DT-19 
                 D-Leu-Met-Tyr-Pro-Thr-Tyr-Deg-Lys-OH 
                 (SEQ ID NO: 15) 
               
               
                 DT-23 
                 D-Phe-Gln-Trp-Aib-Val-Gly-His-Leu-NH 2   
                 (SEQ ID NO: 16) 
               
               
                 DT-24 
                 D-Phe-Gln-Trp-Ala-Val-Aib-His-Leu-NH 2   
                 (SEQ ID NO: 17) 
               
               
                 DT-26 
                 D-Phe-Gln-Trp-Aib-Val-Gly-His-Ile-NH 2   
                 (SEQ ID NO: 18) 
               
               
                 DT-27 
                 D-Phe-Gln-Trp-Ala-Val-Aib-His-Ile-NH 2   
                 (SEQ ID NO: 19) 
               
               
                 DT-31 
                 Aib-Met-Gln-Trp-Phe-Aib-NH 2   
                 (SEQ ID NO: 20) 
               
               
                 DT-33 
                 D-Leu-Met-Gln-Trp-Phe-Aib-NH 2   
                 (SEQ ID NO: 21) 
               
               
                 DT-34 
                 D-Arg-Pro-Lys-Pro-Aib-Gln-D-Trp-Phe-D- 
                 (SEQ ID NO: 22) 
               
               
                   
                 Trp-Aib-Leu-NH 2   
               
               
                 DT-62A 
                 Ala-Aib-Cys-Lys-Asn-Phe-Phe-D-Trp-Lys- 
                 (SEQ ID NO: 23) 
               
               
                   
                 Thr-Phe-Thr-Ser-Cys (3-14 Disulphide 
               
               
                   
                 bond) 
               
               
                 DT-62B 
                 Ala-Aib-Cys-Lys-Asn-Phe-Phe-D-Trp-Lys- 
                 (SEQ ID NO: 24) 
               
               
                   
                 Thr-Phe-Thr-Ser-Cys 
               
               
                 DT-71 
                 D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Aib-Thr- 
                 (SEQ ID NO: 25) 
               
               
                   
                 NH 2   
               
               
                   
               
             
          
         
       
     
     MuJ-7 was tested against primary tumor cells of human colon adenocarcinoma, and each of the peptide analogs comprising MuJ-7 was tested individually against human colon adenocarcinoma cells. A three day MTT cytotoxicity assay was performed as described in U.S. patent application Ser. No. 08/727,679. The percent killing achieved by individual peptides was in the range of 54 to 79% while percent killing achieved by MuJ-7 was 94%. 
     Five different subcombinations of seven peptide analogs comprising MuJ-7 were tested against human colon adenocarcinoma cells. Each subcombination was tested by performing a one day MTT cytotoxicity assay as described in U.S. patent application Ser. No. 08/727,679. The percent killing achieved by the subcombinations was in the range of 64.7% to 94.9%. 
    
    
     EXAMPLE 1 
     ECV.304 cells collected at exponential growth phase were resuspended in medium (3.3×10 6  cells/ml in RPMI 1640 containing 10% FBS). 150 μl of medium was added to the wells of a 96-well tissue culture plate (Nunc, Denmark) followed by 30 μl of cell suspension. The plate was left in incubator (37° C., 5% CO 2 ) overnight. Each of the seven peptides of the combination MuJ-7 were added at three molar concentrations of 10 −7 M, 10 −8 M and 10 −9 M. 20 μl of MuJ-7 at three concentrations of N/10, N and 10N was added to marked wells of the 96-well plate. The value of N for each of the individual peptides was 10 −8 M for VIP 2 , BOM 1 , SP 1 , VIP 3 , and SOM 2  and 10 −7 M for VIP 1 , and 10 −9 M for SOM 1 . Each concentration was plated in triplicate. 20 μl of medium alone was added to control wells while wells without cells served as blanks. A total volume of 200 μl was ensured in each well and the plate was left in an incubator (37° C., 5% CO 2 ). After 72 hours of incubation an MTT assay was performed and percentage inhibition in proliferation of treated cells was calculated with respect to control cells. Results are given in Table I. 
     
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Percent inhibition in proliferation of endothelial cells on treatment with different concentrations of MuJ-7 
               
               
                 and its constituent peptides. 
               
             
          
           
               
                   
                 Percent Inhibition Proliferation (%) 
               
             
          
           
               
                   
                 VIP 2   
                 BOM 1   
                 SP 1   
                 VIP 1   
                 VIP 3   
                 SOM 2   
                 SOM 1   
                 MuJ-7 
               
             
          
           
               
                 Cell line 
                 −7 
                 −8 
                 −9 
                 −7 
                 −8 
                 −9 
                 −7 
                 −8 
                 −9 
                 −7 
                 −8 
                 −9 
                 −7 
                 −8 
                 −9 
                 −7 
                 −8 
                 −9 
                 −7 
                 −8 
                 −9 
                 N/10 
                 N 
                 10N 
               
               
                   
               
             
          
           
               
                 EaHy.926 
                 32 
                 28 
                 42 
                 38 
                 31 
                 32 
                 49 
                 32 
                 37 
                 25 
                 24 
                 23 
                 21 
                 9 
                 24 
                 36 
                 38 
                 25 
                 36 
                 36 
                 22 
                 36 
                 44 
                 38 
               
               
                 ECV.304 
                 24 
                 25 
                 22 
                 21 
                 22 
                 19 
                 18 
                 24 
                 22 
                 15 
                 17 
                 16 
                 21 
                 25 
                 22 
                 20 
                 19 
                 17 
                 20 
                 20 
                 19 
                 24 
                 31 
                 28 
               
               
                   
               
             
          
         
       
     
     EXAMPLE 2 
     ECV304 cells collected at exponential growth phase were resuspended in medium (3.3×10 6  cells/ml in RPMI 1640 containing 10% FBS). 150 μl of medium was added to the wells of a 96-well tissue culture plate (Nunc, Denmark) followed by 30 μl of cell suspension. The plate was left in incubator (37° C., 5% CO 2 ) overnight. Novel peptides shown below were incubated with cells at concentration range of 10 −7  to 10 −9  M (except DT71 at concentration range 10 −8  to 10 −10  M). Each concentration was plated in triplicates. 20 μl of medium alone was added to control wells while wells without cells served as blanks. A total volume of 200 μl was ensured in each well and plate was left in incubator (37° C., 5% CO 2 ). After 72 hours of incubation an MTT assay was performed and percentage inhibition in proliferation of treated cells was calculated with respect to control cells. Results are given in Table II. 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 SEQUENCE LISTING OF NEW PEPTIDE ANALOGS 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 DT-11 
                 Aib-Met-Tyr-Pro-Thr-Tyr-Aib-Lys-OH 
                 (SEQ ID NO: 8) 
               
               
                 DT-12 
                 D-Leu-Met-Tyr-Pro-Thr-Tyr-Aib-Lys-OH 
                 (SEQ ID NO: 9) 
               
               
                 DT-13 
                 Leu-Met-Tyr-Pro-Thr-D-Tyr-Leu-Lys-OH 
                 (SEQ ID NO: 10) 
               
               
                 DT-14 
                 Leu-Met-Tyr-Pro-Thr-Tyr-D-Leu-Lys-OH 
                 (SEQ ID NO: 11) 
               
               
                 DT-15 
                 Leu-Met-D-Tyr-Pro-Thr-Tyr-D-Leu-Lys-OH 
                 (SEQ ID NO: 12) 
               
               
                 DT-16 
                 D-Leu-Met-Tyr-Pro-Thr-Tyr-D-Leu-Lys-OH 
                 (SEQ ID NO: 13) 
               
               
                 DT-18 
                 Aib-Met-Tyr-Pro-Thr-Tyr-Deg-Lys-OH 
                 (SEQ ID NO: 14) 
               
               
                 DT-19 
                 D-Leu-Met-Tyr-Pro-Thr-Tyr-Deg-Lys-OH 
                 (SEQ ID NO: 15) 
               
               
                 DT-23 
                 D-Phe-Gln-Trp-Aib-Val-Gly-His-Leu-NH 2   
                 (SEQ ID NO: 16) 
               
               
                 DT-24 
                 D-Phe-Gln-Trp-Ala-Val-Aib-His-Leu-NH 2   
                 (SEQ ID NO: 17) 
               
               
                 DT-26 
                 D-Phe-Gln-Trp-Aib-Val-Gly-His-Ile-NH 2   
                 (SEQ ID NO: 18) 
               
               
                 DT-27 
                 D-Phe-Gln-Trp-Ala-Val-Aib-His-Ile-NH 2   
                 (SEQ ID NO: 19) 
               
               
                 DT-31 
                 Aib-Met-Gln-Trp-Phe-Aib-NH 2   
                 (SEQ ID NO: 20) 
               
               
                 DT-33 
                 D-Leu-Met-Gln-Trp-Phe-Aib-NH 2   
                 (SEQ ID NO: 21) 
               
               
                 DT-34 
                 D-Arg-Pro-Lys-Aib-Gln-D-Trp-Phe-D-Trp- 
                 (SEQ ID NO: 22) 
               
               
                   
                 Aib-Leu-NH 2   
               
               
                 DT-62A 
                 Ala-Aib-Cys-Lys-Asn-Phe-Phe-D-Trp-Lys- 
                 (SEQ ID NO: 23) 
               
               
                   
                 Thr-Phe-Thr-Ser-Cys (3-14 Disulphide 
               
               
                   
                 bond) 
               
               
                 DT-62B 
                 Ala-Aib-Cys-Lys-Asn-Phe-Phe-D-Trp-Lys- 
                 (SEQ ID NO: 24) 
               
               
                   
                 Thr-Phe-Thr-Ser-Cys 
               
               
                 DT-71 
                 D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Aib-Thr- 
                 (SEQ ID NO: 25) 
               
               
                   
                 NH 2   
               
               
                   
               
             
          
         
       
     
     
       
         
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                 TABLE 2 
               
             
             
               
                   
                   
               
               
                   
                 PERCENT INHIBITION OF PROLIFERATION 
               
             
          
           
               
                   
                 COMPOUND 
                 10 −7 M 
                 10 −8 M 
                 10 −9 M 
               
               
                   
                   
               
             
          
           
               
                   
                 DT-11 
                 16.7 
                 18.7 
                 19.9 
               
               
                   
                 DT-12 
                 20.3 
                 16.1 
                 15.5 
               
               
                   
                 DT-13 
                 22.6 
                 20.3 
                 18.4 
               
               
                   
                 DT-14 
                 17.0 
                 6.0 
                 20.5 
               
               
                   
                 DT-15 
                 20.4 
                 13.2 
                 20.0 
               
               
                   
                 DT-16 
                 24.0 
                 24.7 
                 37.9 
               
               
                   
                 DT-18 
                 23.3 
                 10.9 
                 14.4 
               
               
                   
                 DT-19 
                 11.9 
                 7.3 
                 21.0 
               
               
                   
                 DT-23 
                 3.5 
                 17.2 
                 30.0 
               
               
                   
                 DT-24 
                 6.4 
                 9.8 
                 10.7 
               
               
                   
                 DT-26 
                 0 
                 0 
                 0 
               
               
                   
                 DT-27 
                 14.3 
                 11.9 
                 6.7 
               
               
                   
                 DT-31 
                 1.3 
                 14.1 
                 6.1 
               
               
                   
                 DT-33 
                 8.7 
                 15.8 
                 13.9 
               
               
                   
                 DT-34 
                 12.1 
                 15.6 
                 14.3 
               
               
                   
                 DT-62A 
                 11.7 
                 20.1 
                 25.7 
               
               
                   
                 DT-62B 
                 14.0 
                 3.8 
                 12.8 
               
               
                   
                   
                 10. −8 M 
                 10 −9 M 
                 10 −10 M 
               
               
                   
                 DT-71 
                 14.6 
                 12.1 
                 9.4 
               
               
                   
                   
               
             
          
         
       
     
     EXAMPLE 3 
     Polycarbonate filter transwell inserts (24 well size) with 8 μm pores (Nunc, Denmark) were used for the migration assay. ECV304 (10 4  cells/200 ul DMEM containing 0.1% BSA) was added to the upper chamber. The lower chamber contained 600 ul of DMEM with 0.1% BSA. Individual peptides of the composition MuJ-7 at six different concentrations each of N/100, N/10, N, 10N, 100N and 1000N were added directly to the upper well and the plate incubated at 37° C. for 24 hours. The cells migrated to the lower chamber were viewed randomly at five different phase-contrast microscopic fields and the total number of cells counted using Video Pro 32 Image Analysis system. Percent inhibition in migration was determined with reference to the Control. Similarly, five different concentrations of N/100, N/10, N, 10N, 100N and 1000N of MuJ-7 were added directly to the upper well and the plate incubated at 37° C. for 24 hours. The cells migrated to the lower chamber were counted as described above. The molarity representing N for each peptide is given in Table No. 3. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Percent inhibition in migration of endothelial cells ECV.304 
               
               
                 across a 8μ filter on treatment with various concentrations 
               
               
                 of MuJ-7 and its constituent peptides. 
               
             
          
           
               
                   
                 Percent inhibition in migration (%) 
                   
               
             
          
           
               
                 Peptide 
                 N/100 
                 N/10 
                 N 
                 10N 
                 100N 
                 1000N 
               
               
                   
               
             
          
           
               
                 VIP 2   
                 0.00 
                 91.75 
                 89.05 
                 88.15 
                 92.90 
                 0.00 
               
               
                 BOM 1   
                 0.00 
                 18.25 
                 32.85 
                 24.10 
                 63.95 
                 0.00 
               
               
                 SP 1   
                 13.97 
                 94.10 
                 90.50 
                 92.65 
                 78.10 
                 18.10 
               
               
                 VIP 1   
                 37.53 
                 52.60 
                 69.40 
                 62.60 
                 66.52 
                 0.00 
               
               
                 VIP 3   
                 0.00 
                 32.80 
                 92.45 
                 23.90 
                 11.62 
                 0.00 
               
               
                 SOM 2   
                 7.21 
                 90.70 
                 92.30 
                 89.75 
                 82.15 
                 58.00 
               
               
                 SOM 1   
                 84.27 
                 78.50 
                 88.11 
                 88.75 
                 73.73 
                 25.00 
               
               
                 MuJ-7 
                 0.00 
                 93.39 
                 95.69 
                 97.30 
                 21.6 
                 — 
               
               
                   
               
             
          
         
       
     
     where the value of N for each peptide is 10 −8  M for VIP 2 , BOM 1 , SP 1 , VIP 3 , and SOM 2  and 10 −7 M for VIP 1 , and 10 −9 M for SOM 1 . 
     EXAMPLE 4 
     Polycarbonate filter trasnwell inserts (24 well size) with 8 μm pores (Nuno, Denmark) were used for the migration assay. ECV304 (10 4  cells/200 ul DMEM containing 0.1% BSA) was added to the upper chamber. The lower chamber contained 600 ul of DMEM with 0.1% BSA. Individual peptides at optimum concentration (as indicated in Table 4) were added directly to the upper well and the plate incubatedat 37° C. for 24 hours. The sequence of each of the peptides added are described above. The cells migrated to the lower chamber were viewed randomly at five different phase-contrast microscopic fields and the total number of cells counted using Video Pro 32 Image Analysis system. Percent inhibition in migration was determined with reference to Control. 
     
       
         
               
             
               
               
               
             
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Percent inhibition in migration of endothelial cells ECV304 
               
               
                 across a 8μ filter on treatment with analogs 
               
             
          
           
               
                   
                   
                 PERCENT 
               
               
                   
                   
                 INHIBITION 
               
               
                   
                 COMPOUND 
                 MIGRATION 
               
               
                   
                   
               
             
          
           
               
                   
                 DT-11(−8M) 
                 82.5 
               
               
                   
                 DT-12 (−8M) 
                 58.7 
               
               
                   
                 DT-13 (−8M) 
                 89.7 
               
               
                   
                 DT-14 (−8M) 
                 77.3 
               
               
                   
                 DT-15 (−8M) 
                 73.2 
               
               
                   
                 DT-16 (−8M) 
                 90.07 
               
               
                   
                 DT-18 (−8M) 
                 73.2 
               
               
                   
                 DT-19 (−8M) 
                 80.4 
               
               
                   
                 DT-23 (−8M) 
                 0 
               
               
                   
                 DT-24 (−8M) 
                 0 
               
               
                   
                 DT-26 (−8M) 
                 0 
               
               
                   
                 DT-27 (−8M) 
                 0 
               
               
                   
                 DT-31 (−8M) 
                 0 
               
               
                   
                 DT-33 (−8M) 
                 41.4 
               
               
                   
                 DT-34 (−8M) 
                 53.6 
               
               
                   
                 DT-62A (−8M) 
                 62.2 
               
               
                   
                 DT-62B (−8M) 
                 8.5 
               
               
                   
                 DT-71 (−9M) 
                 68.2 
               
               
                   
                   
               
             
          
         
       
     
     EXAMPLE 5 
     Polycarbonate filter transwell inserts (24 well size) with 8 μm pores (Nunc, Denmark) were used for the migration assay as described in the previous example. ECV304 (10 4  cells/200 ul DMEM containing 0.1% BSA) was added to the upper chamber. 
     The lower chamber contained 600 ul of DMEM with 0.1% BSA. Ten different subcombinations of MuJ-7 listed in Table 5 were tested for inhibition of migration of endothelial cells across the filter as described previously. None of the ten subcombinations caused percent inhibition of migration greater than MuJ-7 and was in the range of 11.3 to 69.3% (Table 6). 
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Ten combinations of MuJ-7 with different ratios of its constituent peptides. 
               
             
          
           
               
                 Combination 
                 VIP 1   
                 VIP 2   
                 VIP 3   
                 SOM 1   
                 SOM 2   
                 BOM 1   
                 SP 1   
               
               
                   
               
             
          
           
               
                 MuJ-7 
                 −7 
                 −8 
                 −8 
                 −9 
                 −8 
                 −8 
                 −8 
               
               
                 1 
                 −8 
                 −8 
                 −8 
                 −9 
                 −8 
                 −8 
                 −6 
               
               
                 2 
                 −6 
                 −8 
                 −8 
                 −9 
                 −8 
                 −8 
                 −10 
               
               
                 3 
                 −7 
                 −9 
                 −8 
                 −9 
                 −8 
                 −6 
                 −8 
               
               
                 4 
                 −7 
                 −7 
                 −8 
                 −9 
                 −8 
                 −10 
                 −8 
               
               
                 5 
                 −7 
                 −6 
                 −7 
                 −9 
                 −8 
                 −8 
                 −8 
               
               
                 6 
                 −7 
                 −10 
                 −9 
                 −9 
                 −8 
                 −8 
                 −8 
               
               
                 7 
                 −5 
                 −8 
                 −8 
                 −9 
                 −8 
                 −8 
                 −7 
               
               
                 8 
                 −9 
                 −8 
                 −8 
                 −9 
                 −8 
                 −8 
                 −9 
               
               
                 9 
                 −7 
                 −8 
                 −6 
                 −9 
                 −8 
                 −7 
                 −8 
               
               
                 10  
                 −7 
                 −8 
                 −10 
                 −9 
                 −8 
                 −9 
                 −8 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Percent inhibition in migration of endothelial cells ECV.304 
               
               
                 across a 8μ filter on treatment with ten different 
               
               
                 combinations of MuJ-7 
               
             
          
           
               
                   
                 Subcombination 
                 % Inhibition Migration 
               
               
                   
                   
               
               
                   
                 MuJ-7 
                 95.7 
               
               
                   
                 1 
                 38.6 
               
               
                   
                 2 
                 69.3 
               
               
                   
                 3 
                 19.3 
               
               
                   
                 4 
                 50.0 
               
               
                   
                 5 
                 11.3 
               
               
                   
                 6 
                 28.4 
               
               
                   
                 7 
                 13.6 
               
               
                   
                 8 
                 37.5 
               
               
                   
                 9 
                 32.9 
               
               
                   
                 10  
                 25.0 
               
               
                   
                   
               
             
          
         
       
     
     EXAMPLE 6 
     Matrigel (350 ul) was placed into each well of a 24-well culture plate at 4° C. and was allowed to polymerize by incubation at 37° C. for 30 min. ECV304 (1.5×10 4 ) were seeded on the Matrigel in 500 ul DMEM supplemented with 10% FBS. MuJ-7 at four different concentrations of 0.25×, 0.5×, ×, &amp; 5.0× were added and. the plate incubated at 37° C. for 24 hour. “×” denotes normal concentration of MuJ-7 where molar concentration of individual peptides was 10 −8  M for VIP 2 , BOM 1 , SP 1 , VIP 3  and SOM 2  and 10 −7  M for VIP 1  and 10 −9  M for SOM 1 . Tube like structures formed, the area of which was individually counted at five different phase-contrast microscopic fields using Video Pro 32 Image Analysis system. Percent inhibition in tube area was determined with reference to untreated cells. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Percent inhibition in Tube-like-structure (TLS) area on treatment 
               
               
                 with various concentrations of MuJ-7. 
               
             
          
           
               
                   
                 Concentration of MuJ-7 
                 Percent in tube area 
               
               
                   
                   
               
               
                   
                  0.25x 
                 28.4 
               
               
                   
                 0.5x 
                 39.3 
               
               
                   
                 x 
                 47.8 
               
               
                   
                 5.0x 
                 58.1 
               
               
                   
                   
               
             
          
         
       
     
     EXAMPLE 7 
     Matrigel (350 ul) was placed into each well of a 24-well culture plate at 4° C. and was allowed to polymerize by incubation at 37° C. for 30 min. ECV304 (1.5×10 4 ) were seeded on the Matrigel in 500 ul DMEM supplemented with 10% FBS. New analogs of peptides as shown in FIG. 1 were added at optimum concentration as denoted in Table 8, and the plate incubated at 37° C. for 24 hours. Tube like structures formed, the area of which was individually counted at five different phase-contrast microscopic fields using Video Pro 32 Image Analysis system. Percent inhibition in tube length was determined with reference to untreated cells. 
     
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                   
                   
                   
                 TUBE 
                 INHIBITION OF 
               
               
                 PEPTIDE 
                 SPROUTING 
                 INTUSSUSCEPTION 
                 FORMATION 
                 TUBE LENGTH 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 DT-10 (−8M) 
                 + 
                 + 
                 + 
                 59.5 
               
               
                 DT-11 (−8M) 
                 ++ 
                 + 
                 + 
                 3.6 
               
               
                 DT-12 (−8M) 
                 ++ 
                 + 
                 ++ 
                 17.1 
               
               
                 DT-13 (−8M) 
                 ++ 
                 ++ 
                 ++ 
                 16.2 
               
               
                 DT-14 (−8M) 
                 ++ 
                 + 
                 + 
                 24.3 
               
               
                 DT-15 (−8M) 
                 ++ 
                 + 
                 ++ 
                 0 
               
               
                 DT-16 (−8M) 
                 ++ 
                 + 
                 + 
                 18.9 
               
               
                 DT-18 (−8M) 
                 ++ 
                 ++ 
                 ++ 
                 0 
               
               
                 DT-19 (−8M) 
                 ++ 
                 + 
                 + 
                 37.8 
               
               
                 DT-23 (−8M) 
                 + 
                 + 
                 + 
                 53.1 
               
               
                 DT-24 (−8M) 
                 ++ 
                 + 
                 + 
                 16.2 
               
               
                 DT-26 (−8M) 
                 ++ 
                 ++ 
                 ++ 
                 0 
               
               
                 DT-27 (−8M) 
                 + 
                 + 
                 + 
                 29.7 
               
               
                 DT-31 (−8M) 
                 + 
                 + 
                 + 
                 17.1 
               
               
                 DT-33 (−8M) 
                 ++ 
                 ++ 
                 ++ 
                 13.5 
               
               
                 DT-34 (−8M) 
                 ++ 
                 ++ 
                 ++ 
                 0 
               
               
                 DT-62A (−8M) 
                 ++ 
                 + 
                 ++ 
                 44.1 
               
               
                 DT-62B (−8M) 
                 ++ 
                 ++ 
                 ++ 
                 0 
               
               
                 DT-71 (−9M) 
                 ++ 
                 + 
                 ++ 
                 9.9 
               
               
                   
               
             
          
         
       
     
     EXAMPLE 8 
     Matrigel (350 ul) was placed into each well of a 24-well culture plate at 4° C. and was allowed to polymerize by incubation at 37° C. for 30 min. ECV304 (1.5×10 4 ) were seeded on the Matrigel in each of the 24 wells. 500 ul DMEM supplemented with 10% FBS and was added to 8 wells. These wells served as controls. Culture supernatants of primary tumor cells of human colon adenocacinoma from a confluent culture in log phase of growth was filtered through a 0.22 u filter and its pH adjusted to 7.4 with sodium bicarbonate. 500 ul of this supernatant was added to another 8 wells, containing ECV304 (1.5×10 4 ) cells seeded on Matrigel. To the remaining 8 wells, MuJ-7 was added and incubated at 37° C. for 24 hours. Five different phase-contrast microscopic fields (4×) were reviewed and total tube length of the tube-like-structures (TLS) measured using Video Pro 32 Image Analysis system. Percent inhibition of TLS was calculated with reference to Controls. In controls, although sprouting was visible at day 1, intussusception was still missing. However, endothelial cells treated PTC culture supernatant showed enhanced sprouting as well as intussusception leading to completion of tube formation. This clearly suggests pro-angiogenic activity of factors in culture supernatant of primary tumor cells of human colon adenocarcinoma. Treating PTC culture supernatant treated endothelial cells with MuJ-7 resulted in complete inhibition of intussusception and a significant decrease in sprouting. This indicates the ability of MuJ-7 to inhibit two key steps of a new blood vessel formation, i. e., sprouting and angiogenesis. A qualitative representation of the effect is given in Table 9. 
     
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 Qualitative representation of the effect of MuJ-7 on 
               
               
                 Tube-like-structure activity of PTC culture supernatant stimulated 
               
               
                 endothelial cells. 
               
             
          
           
               
                   
                   
                 PTC culture 
                   
               
               
                   
                 Untreated 
                 supernatant stimulated 
                 MuJ-7 treated 
               
               
                   
                   
               
             
          
           
               
                 Sprouting 
                 ++ 
                 +++ 
                 + 
               
               
                 Intussusception 
                 + 
                 ++ 
                 − 
               
               
                 Tube formation 
                 + 
                 ++ 
                 − 
               
               
                   
               
             
          
         
       
     
     EXAMPLE 9 
     Matrigel (350 ul) was placed into each well of a 24-well culture plate at 4° C. and was allowed to polymerize by incubation at 37° C. for 30 min. ECV304 (1.5×10 4 ) were seeded on the Matrigel in 500 ul DMEM supplemented with 10% FBS. Individual peptides of the composition MuJ-7 at three different concentrations each of N/10, N, and 10N were added and the plate incubated at 37° C. for 24 hours. Tube like structures formed, the length of which were individually counted at five different phase-contrast microscopic fields using Video Pro 32 Image Analysis system. Percent inhibition in tube length was determined with reference to Control. Similarly, three different concentrations of N/10, N, and 10N of MuJ-7 were added to the well and the plate incubated at 37° C. for 24 hours. Percent inhibition in tube length was determined with reference to Control as described previously (Table 10). 
     
       
         
               
             
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 10 
               
             
             
               
                   
               
               
                 Percent inhibition in Tube-like-structure (TLS) length on 
               
               
                 treatment with various concentrations of MuJ-7 and its 
               
               
                 constituent peptides. 
               
             
          
           
               
                   
                 Percent inhibition in TLS activity (%) 
               
             
          
           
               
                   
                 Peptide 
                 N/10 
                 N 
                 10N 
               
               
                   
                   
               
             
          
           
               
                   
                 VIP 2   
                 0.0 
                 1.0 
                 3.3 
               
               
                   
                 BOM 1   
                 0.0 
                 0.0 
                 2.9 
               
               
                   
                 SP 1   
                 14.4 
                 8.9 
                 3.0 
               
               
                   
                 VIP 1   
                 14.9 
                 11.7 
                 13.1 
               
               
                   
                 VIP 1   
                 8.5 
                 22.1 
                 17.9 
               
               
                   
                 SOM 2   
                 1.0 
                 16.5 
                 16.8 
               
               
                   
                 SOM 2   
                 7.0 
                 15.1 
                 16.7 
               
               
                   
                 MuJ-7 
                 6.9 
                 15.6 
                 22.27 
               
               
                   
                   
               
             
          
         
       
     
     where the value of N for each peptide is 10 −8 M for VIP 2 , BOM 1 , SP 1 , VIP 3 , and SOM 2  and 10 −7 M for VIP 1 , and 10 −9 M for SOM 1 . 
     EXAMPLE 10 
     Matrigel (350 ul) was placed into each well of a 24-well culture plate at 4° C. and was allowed to polymerize by incubation at 37° C. for 30 min. ECV304 (1.5×10 4 ) were seeded on the Matrigel in each of the 24 wells. 500 ul DMEM supplemented with 10% FBS was added to 8 wells. These wells served as controls. Culture supenatants of primary tumor cells of human colon adenocarcinoma from a confluent culture in log phase of growth was filtered through a 0.22 u filter and its pH adjusted to 7.4 with sodium bicarbonate. 500 ul of this supematant was added to another 8 wells contains ECV304 (1.5×10 4 ) cells seeded on Matrigel. To the remains 8 wells, MuJ-7 was added and incubated at 37° C. for 24 hours. Five different phase-contrast microscopic fields (4×) were viewed and total tube length of the tube-like-structures (TLS) measured using Video Pro 32 Image Analysis system. Percent inhibition of TLS was calculated with reference to Controls. In controls, 76.9% of TLS were in the range of 50-69 μm and remaining were between 69-107 μm. There were no structures greater than 107 μm. In PTC stimulated wells, the percentage of TLS in range of 50-69 μm reduced to 50.0% while 27.3% were in the range of 69-107 μm. It is interesting to note that there were approximately 18.0% TLS in the range of 107-183 μm and 4.55% TLS in 221-240 μm. Hence there was a significant increase in the length of TLS in PTC supernatant stimulated cells as compared to the unstimulated controls. This clearly suggests pro-angiogenic activity of factors in culture supematant of primary tumor cells of human colon adenocarcinoma. In the third group of wells, which were treated with MuJ-7, 81.25% TLS were in the range of 50-69 μm and 18.75% in the range of 69-88 μm (Table 6). This reversal of TLS activity to control level on treatment with MuJ-7 clearly suggests anti angiogenic activity of MuJ-7. 
     
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 11 
               
             
             
               
                   
               
               
                 Percent inhibition in TLS length of PTC culture supernatant 
               
               
                 endothelial cells treated with MuJ-7. 
               
             
          
           
               
                 Tube length Range 
                   
                 PTC culture 
                   
               
               
                 (in microns) 
                 Controls 
                 supernatant stimulated 
                 MuJ-7 treated 
               
               
                   
               
             
          
           
               
                 50-69 
                 76.92 
                 50.00 
                 81.25 
               
               
                 69-88 
                 7.69 
                 4.55 
                 18.75 
               
               
                  88-107 
                 15.38 
                 22.73 
                 0.00 
               
               
                 107-126 
                 0.00 
                 4.55 
                 0.00 
               
               
                 126-145 
                 0.00 
                 9.09 
                 0.00 
               
               
                 145-164 
                 0.00 
                 0.00 
                 0.00 
               
               
                 164-183 
                 0.00 
                 4.55 
                 0.00 
               
               
                 183-202 
                 0.00 
                 0.00 
                 0.00 
               
               
                 202-221 
                 0.00 
                 0.00 
                 0.00 
               
               
                 221-240 
                 0.00 
                 4.55 
                 0.00 
               
               
                   
               
             
          
         
       
     
     REFERENCES 
     1. Cancer: Principles &amp; Practice of Oncology, Fifth Edition, edited by Vincent T. DeVita, Jr., Samuel Hellinan, Steven A. Rosenberg. Lippincott-Raven Publishers, Philadelphia © 1997. 
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