METHODS OF INCREASING SENSITIVITY OF AN INDIVIDUAL TO OB PROTEIN BY UPREGULATING OB PROTEIN RECEPTOR

Abstract: Disclosed are methods for increasing the sensitivity of functional OB protein receptor and uses thereof.

Description: METHODS OF INCREASING SENSITIVITY OF AN INDIVIDUAL TO OB PROTEIN BY UPREGULATING OB PROTEIN RECEPTOR FIELD OF THE INVENTION The present invention relates to methods of reducing amounts of OB protein compositions administered for desired therapeutic or cosmetic effect through the use of agents which increase the sensitivity to OB protein or the affinity or availability of functional OB protein receptor . BACKGROUND Although the molecular basis for obesity is largely unknown, the identification of the "OB gene" and protein encoded ("OB protein" or "leptin") has shed some light on mechanisms the body uses to regulate body fat deposition. See, PCT publication, WO 96/05309 (12/22/96), Friedman et al . ; Zhang et al . , Nature 372 : 425-432 (1994) ; see also, the Correction at Nature 374: 479 (1995) . The OB protein is active in vivo in both ob/ob mutant mice (mice obese due to a defect in the production of the OB gene product) as well as in normal, wild type mice. The biological activity manifests itself in, among other things, weight loss. See generally, Barinaga, "Obese" Protein Slims Mice, Science 269: 475-476 (1995) . The OB protein, derivatives and use thereof as modulators for the control of weight and adiposity of animals, including mammals and humans, has been disclosed in greater detail in PCT publication WO 96/05309 (12/22/96), hereby incorporated by reference, including figures . The other biological effects of OB protein are not well characterized. It is known, for instance, that in ob/ob mutant mice, administration of OB protein results in a decrease in serum insulin levels, and serum glucose levels. It is also known that administration of OB protein results in a decrease in body f t. This was observed in both ob/ob mutant mice, as well as non-obese normal mice. Pelleymounter et al . , Science 269 : 540-543 (1995); Halaas et al . , Science 169.: 543-546 (1995). See also, Campfield et al . , Science 269 : 546-549 (1995) (Peripheral and central administration of microgram doses of OB protein reduced food intake and body weight of ob/ob and diet-induced obese mice but not in db/db obese mice.) In none of these reports have toxicities been observed, even at the highest doses. There are other agents which may play a role in the regulation of insulin metabolism. Thiazolidinediones are a class of antihyperglycemic agents with an as yet unknown mechanism of action. See Wilson et al . , J. Med. Chem., 3£: 665-668 (1996); Kallen et al., P.N.A.S., 93 . : 5793-5796 (1996); Sohda et al . , Chem. Pharm. Bull., 43.: 2168-2172 (1995); Swanson et al., Drug Dev. Res., 3_5_: 69-82 (1995); Kletzien et al . , Mol. Pharmacol., __2 : 558-62 (1992). It has been hypothesized that this class of drugs acts to reduce hyperglycemia by 1) increasing insulin sensitivity or 2) by activating the transcription factor , peroxisome proliferator-activated receptor (PPAR) (gamma) . PPAR (gamma) is thought to play a role in the conversion of primary fibroblasts to adipocytes . See Wilson et al., J. Med. Chem., 39.: 665-668 (1996); Kallen et al . , P.N.A.S., 92: 5793-5796 (1996); Swanson et al . , Drug Dev. Res., 35 . : 69-82 (1995). Recently it has been reported that one of the thiazolidinediones (AD-5057) was able to significantly reduce adipose leptin mRNA and serum leptin levels in obese rats and mice without substantial effect on food intake, with minor elevations in body weight. Zhang et al . , J. Biol. Chem., 271 : 9455-9459 (1996) . Another report indicated that BRL496553, another type of thiazolidinedione, also reduced leptin mRNA in adipocytes in vitro . Kallen et al., P.N.A.S., 93.: 5793-5796 (1996). Thus, thiazolidinediones are thought to decrease endogenous OB protein. Although thiazolidinediones are thought to increase insulin sensitivity, there are currently no known agents which increase an individual's sensitivity to OB protein (or leptin) . Such increased sensitivity would be advantageous to leptin consumers, as increased sensitivity may contribute to lower doses required or less frequent dosing. SUMMARY OF THE INVENTION The present invention is based on the hypothesis that thiazolidinedione compositions provide for increased sensitivity to OB protein. Although not wishing to be bound by theory, it is possible that thiazolidinedione compositions "upregulate" OB protein receptor available for signal transduction, i.e., increase the number of OB receptor, or increase the affinity of OB receptor for its ligand (OB protein) . This increase in the availability or affinity of functional OB protein receptor provides for increased signal transduction of endogenous and/or exogenous OB protein. Surprisingly, and importantly, this provides means for reducing the amount and/or frequency of dosages of exogenous OB protein for therapeutic and/or cosmetic purposes. Therefore, in one aspect, the present invention provides methods of weight modulation and/or fat deposition in an individual by the administration of one or more compositions which function to increase the availability or affinity of functional OB protein receptors in an individual, such as thiazolidinedione compositions. In another aspect, the present invention provides for methods of weight modulation and/or fat deposition in an individual by the administration of one or more compositions which function to increase the availability of functional OB protein receptors, such as a thiazolidinedione composition, in an individual, in combination with administration of an OB protein. In yet another aspect, the present invention provides for methods to reduce the amount and/or frequency of dosage of exogenous an OB protein by administration of a composition which functions to increase the availability of functional OB protein receptors, such as a thiazolidinedione composition. In yet other aspects , the present invention provides for methods for the treatment of co-morbidities associated with excess fat, such as diabetes, dys- or hyperlipidemias, lack of fertility, and also potentially an increase in insulin sensitivity and/or and increase in lean tissue mass. Related compositions are also provided . BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 : Recombinant urine met OB (double stranded) DNA (SEQ ID NOS: 1 and 2) and amino acid sequence (SEQ ID NO: 3). Figure 2 : Recombinant human met OB analog (double stranded) DNA (SEQ ID NOS: 4 and 5) and amino acid sequence (SEQ ID NO: 6). DETAILED DESCRIPTION Compositions The composition having the ability to increase the sensitivity to OB protein or the availability of functional OB protein receptor in an individual may be selected from among various thiazolidinedione compositions, such as: 2 , 4-thiazolidinediones; opt. substituted thiazolidinediones; 5- [4- [2- (5-methyl-2- phenyl-4-oxazolyl) -2-hydroxyethoxy] enzyl] -2, 4- thiazolidinedione (AD-5070) ; clofibrate; ciglitazone; englitazone; pioglitazone; BRL 49653; troglitazone; M16209; oxazolidinediones; as well as derivatives, analogs, tautomers, enantiomerε, diastereomers , epimers, salts, solvates, esters, prodrugs and metabolites of thiazolidinediones or the compounds above. The OB protein may be selected from recombinant urine set forth below (SEQ. ID No. 3 of Figure 1) , or recombinant human protein as set forth in Zhang et al . , Nature , supra , herein incorporated by reference) or those lacking a glutaminyl residue at position 28. (See Zhang et al, Nature, supra , at page 428.) One may also use the recombinant human OB protein analog as set forth in SEQ. ID. O. 6 of Figure 2, which contains 1) an arginine in place of lysine at position 35 and 2) a leucine in place of isoleucine at position 74. (A shorthand abbreviation for this analog is the recombinant human R->K^5 / ->I^^) . The amino acid sequences for the recombinant human analog and recombinant murine proteins are set forth below with a methionyl residue at the -1 position, however, as with any of the present OB proteins and analogs , the methionyl residue may be absent. The murine protein is substantially homologous to the human protein, particularly as a mature protein, and, further, particularly at the N-terminus. One may prepare an analog of the recombinant human protein by altering (such as substituting amino acid residues) , in the recombinant human sequence, the amino acids which diverge from the murine sequence. Because the recombinant human protein has biological activity in mice, such analog would likely be active in humans. For example, using a human protein having a lysine at residue 35 and an isoleucine at residue 74 according to the numbering of SEQ. ID NO. 6, wherein the first amino acid is valine, and the amino acid at position 146 is cysteine, one may substitute with another amino acid one or more of the amino acids at positions 32, 35, 50, 64, 68, 71, 74, 77, 89, 97, 100, 105, 106, 107, 108, 111, 118, 136, 138, 142, and 145. One may select the amino acid at the corresponding position of the murine protein, (SEQ. ID. NO. 3), or another amino acid. One may further prepare "consensus" molecules based on the rat OB protein sequence. Murakami et al . , Biochem. Biophys. Res. Comm. 209: 944-952 (1995) herein incorporated by reference. Rat OB protein differs from human OB protein at the following positions (using the numbering of SEQ. ID. NO. 6): 4, 32, 33, 3_5, 5 . 0 . , 68, 21, Ik, TL, 78, ££- 92- 1-2Q. 101, 102, 1_5, 106- 107, 108, 111, 118, 136, 138 and 145. One may substitute with another amino acid one or more of the amino acids at these divergent positions. The positions in bold print are those which in which the murine OB protein as well as the rat OB protein are divergent from the human OB protein, and thus, are particularly suitable for alteration. At one or more of these positions, one may substitute an amino acid from the corresponding rat OB protein, or another amino acid. The positions from both rat and murine OB protein which diverge from the mature human OB protein are: 4, 32, 33, 35, 50, 64, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 118, 136, 138, 142, and 145. A human OB protein according to SEQ. ID. NO. 6 (with lysine at position 35 and isoleucine at position 74) having one or more of the above amino acids deleted or replaced with another amino acid, such as the amino acid found in the corresponding rat or murine sequence, may also be effective. In addition, the amino acids found in rhesus monkey OB protein which diverge from the mature human OB protein are (with identities noted in parentheses in one letter amino acid abbreviation): 8 (S) , 35 (R) , 48 (V) , 53(Q), 60(1), 66(1), 67 (N) , 68 ( (L) , 89 (L) , 100 (L) , 108(E), 112 (D) , and 118 (L) . Since the recombinant human OB protein is active in cynomolgus monkeys, a human OB protein according to SEQ. ID. NO. 6 (with lysine at position 35 and isoleucine at position 74) having one or more of the rhesus monkey divergent amino acids replaced with another amino acid, such as the amino acids in parentheses, may be effective. It should be noted that certain rhesus divergent amino acids are also those found in the above murine species (positions 35, 68, 89, 100 and 112). Thus, one may prepare a murine/rhesus/human consensus molecule having (using the numbering of SEQ. ID. NO. 6 having a lysine at position 35 and an isoleucine at position 74) having one or more of the amino acids at positions replaced by another amino acid: 4, 8, 32, 33, 3_5, 48, 50, 53, 60, 64, 66, 67, 68 , 71, 74, 77, 78, . 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145. Other analogs may be prepared by deleting a part of the protein amino acid sequence. For example, the mature protein lacks a leader sequence (-22 to -1) . One may prepare the following truncated forms of human OB protein molecules (using the numbering of SEQ. ID. NO. 6) : (a) amino acids 98-146 (b) amino acids 1-32 (c) amino acids 40-116 (d) amino acids 1-99 and (connected to) 112-146 (e) amino acids 1-99 and (connected to) 112-146 having one or more of amino acids 100-111 placed between amino acids 99 and 112. In addition, the truncated forms may also have altered one or more of the amino acids which are divergent (in the rhesus, rat or murine OB protein) from human OB protein. Furthermore, any alterations may be in the form of altered amino acids, such as peptidomimetics or D-amino acids. Derivatives The present protein (herein the term "protein" is used to include "peptide" and OB analogs, such as those recited infra, unless otherwise indicated) may also be derivatized by the attachment of one or more chemical moieties to the protein moiety. The chemically modified derivatives may be further formulated for intraarterial , intraperitoneal, intramuscular, subcutaneous, intravenous, oral, nasal, pulmonary, topical or other routes of administration. Chemical modification of biologically active proteins has been found to provide additional advantages under certain circumstances, such as increasing the stability and circulation time of the therapeutic protein and decreasing immunogenicity . See U.S. Patent No. 4,179,337, Davis et al . , issued December 18, 1979. For a review, see Abuchowski et al., in Enzymes as Drugs. (J.S. Holcerberg and J. Roberts, eds . pp. 367-383 (1981)). A review article describing protein modification and fusion proteins is Francis, Focus on Growth Factors 3_ : 4-10 (May 1992) (published by Mediscript, Mountview Court, Friern Barnet Lane, London N20, OLD, UK) . The chemical moieties suitable for derivatization may be selected from among various water soluble polymers . The polymer selected should be water soluble so that the protein to which it is attached does not precipitate in an aqueous environment, such as a physiological environment. Preferably, for therapeutic use of the end-product preparation, the polymer will be pharmaceutically acceptable. One skilled in the art will be able to select the desired polymer based on such considerations as whether the polymer/protein conjugate will be used therapeutically, and if so, the desired dosage, circulation time, resistance to proteolysis, and other considerations . For the present proteins and peptides, the effectiveness of the derivatization may be ascertained by administering the derivative, in the desired form (i.e., by osmotic pump, or, more preferably, by injection or infusion, or, further formulated for oral, pulmonary or nasal delivery, for example) , and observing biological effects as described herein . The water soluble polymer may be selected from the group consisting of, for example, polyethylene glycol, copolymers of ethylene glycol/propylene glycol , carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrolidone, poly-1, 3-dioxolane, poly-1, 3 , 6-trioxane, ethylene/maleic anhydride copolymer, polyaminoacids (either homopolymers or random or non-random copolymers) , and dextran or poly(n-vinyl pyrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols, polystyrenemaleate and polyvinyl alcohol. Polyethylene glycol propionaldenhyde may have advantages in manufacturing due to its stability in water. Fusion proteins may be prepared by attaching polyaminoacids to the OB protein (or analog) moiety. For example, the polyamino acid may be a carrier protein which serves to increase the circulation half life of the protein. For the present therapeutic or cosmetic purposes, such polyamino acid should be those which have do not create neutralizing antigenic response, or other adverse response. Such polyamino acid may be selected from the group consisting of serum album (such as human serum albumin) , an antibody or portion thereof (such as an antibody constant region, sometimes called "F c ") or other polyamino acids. As indicated below, the location of attachment of the polyamino acid may be at the N- terminus of the OB protein moiety, C-terminus, or other place, and also may be connected by a chemical "linker" moiety to the OB protein. The polymer may be of any molecular weight, and may be branched or unbranched. For polyethylene glycol, the preferred molecular weight is between about 2 kDa and about 100 kDa (the term "about" indicating that in preparations of polyethylene glycol, some molecules will weigh more, some less, than the stated molecular weight) for ease in handling and manufacturing. Other sizes may be used, depending on the desired therapeutic profile (e.g., the duration of sustained release desired, the effects, if any on biological activity, the ease in handling, the degree or lack of antigenicity and other known effects of the polyethylene glycol to a therapeutic protein or analog) . The number of polymer molecules so attached may vary, and one skilled in the art will be able to ascertain the effect on function. One may mono-derivatize, or may provide for a di-, tri-, tetra- or some combination of derivatization, with the same or different chemical moieties (e.g., polymers, such as different weights of polyethylene glycolε) . The proportion of polymer molecules to protein (or peptide) molecules will vary, as will their concentrations in the reaction mixture. In general, the optimum ratio (in terms of efficiency of reaction in that there is no excess unreacted protein or polymer) will be determined by factors such as the desired degree of derivatization (e.g., mono, di-, tri-, etc.), the molecular weight of the polymer selected, whether the polymer is branched or unbranched, and the reaction conditions. The chemical moieties should be attached to the protein with consideration of effects on functional or antigenic domains of the protein. There are a number of attachment methods available to those skilled in the art. E. . , EP 0 401 384 herein incorporated by reference (coupling PEG to G-CSF) , see also Malik et al . , Exp. Hematol. 20.: 1028-1035 (1992) (reporting pegylation of GM-CSF using tresyl chloride) . For example, polyethylene glycol may be covalently bound through amino acid residues via a reactive group, such as, a free amino or carboxyl group. Reactive groups are those to which an activated polyethylene glycol molecule may be bound. The amino acid residues having a free amino group may include lysine residues and the N-terminal amino acid residue. Those having a free carboxyl group may include aspartic acid residues, glutamic acid residues, and the C-terminal amino acid residue. Sulfhydrl groups may also be used as a reactive group for attaching the polyethylene glycol molecule (s). Preferred for therapeutic purposes is attachment at an amino group, such as attachment at the N-terminus or lysine group. Attachment at residues important for receptor binding should be avoided if receptor binding is desired. One may specifically desire N-terminally chemically modified protein. Using polyethylene glycol as an illustration of the present compositions, one may select from a variety of polyethylene glycol molecules (by molecular weight, branching, etc.), the proportion of polyethylene glycol molecules to protein molecules in the reaction mix, the type of pegylation reaction to be performed, and the method of obtaining the selected N-terminally pegylated protein. The method of obtaining the N-terminally pegylated preparation (i.e., separating this moiety from other monopegylated moieties if necessary) may be by purification of the N-terminally pegylated material from a population of pegylated protein molecules. Selective N-terminal chemical modification may be accomplished by reductive alkylation which exploits differential reactivity of different types of primary amino groups (lysine versus the N-terminal) available for derivatization in a particular protein. Under the appropriate reaction conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group containing polymer is achieved. For example, one may selectively N- terminally pegylate the protein by performing the reaction at a pH which allows one to take advantage of the pK a differences between the e-amino group of the lysine residues and that of the a-amino group of the N-terminal residue of the protein. By such selective derivatization, attachment of a water soluble polymer to a protein is controlled: the conjugation with the polymer takes place predominantly at the N-terminus of the protein and no significant modification of other reactive groups, such as the lysine side chain amino groups, occurs. Using reductive alkylation, the water soluble polymer may be of the type described above, and should have a single reactive aldehyde for coupling to the protein. Polyethylene glycol propionaldehyde, containing a single reactive aldehyde, may be used. An N-terminally monopegylated derivative is preferred for ease in production of a therapeutic. N-terminal pegylation ensures a homogenous product as characterization of the product is simplified relative to di-, tri- or other multi pegylated products. The use of the above reductive alkylation process for preparation of an N-terminal product is preferred for ease in commercial manufacturing. Complexes The OB protein, analog or derivative may be administered complexed to a binding composition. Such binding composition may have the effect of prolonging the circulation time of the OB protein, analog or derivative. Such composition may be a protein (or synonymously, peptide) . An example of a binding protein is OB protein receptor or portion thereof, such as a soluble portion thereof . Other binding proteins may be ascertained by examining OB protein in serum, or by empirically screening for the presence of binding. Such binding will typically not interfere with the ability of OB protein or analog or derivative to bind to endogenous OB protein receptor and/or effect signal transduction. Pharmaceutical Compositions In yet another aspect of the present invention, provided are methods of using pharmaceutical compositions of the proteins, and derivatives. Such pharmaceutical compositions may be for administration by injection, or for oral, pulmonary, nasal, transdermal or other forms of administration. In general, comprehended by the invention are pharmaceutical compositions comprising effective amounts of protein or derivative products of the invention together with pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions include diluents of various buffer content (e.g., Tris-HCl, acetate, phosphate), pH and ionic strength; additives such as detergents and solubilizing agents (e.g., Tween 80, Polysorbate 80), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite) , preservatives (e.g., Thimersol, benzyl alcohol) and bulking substances (e.g., lactose, mannitol) ; incorporation of the material into particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, etc. or into liposomeε. Hylauronic acid may also be used, and this may have the effect of promoting sustained duration in the circulation. Such compositions may influence the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the present proteins and derivatives. See, e.g. , Remington's Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, PA 18042) pages 1435-1712 which are herein incorporated by reference. The compositions may be prepared in liquid form, or may be in dried powder, such as lyophilized form. Implantable sustained release formulations are also contemplated, as are transdermal formulations. Contemplated for use herein are oral solid dosage forms, which are described generally in Remington's Pharmaceutical Sciences, 18th Ed. 1990 (Mack Publishing Co. Easton PA 18042) at Chapter 89, which is herein incorporated by reference. Solid dosage forms include tablets, capsules, pills, troches or lozenges; cachets or pellets. Also, liposomal or proteinoid encapsulation may be used to formulate the present compositions (as, for example, proteinoid microspheres reported in U.S. Patent No. 4,925,673). Liposomal encapsulation may be used and the liposomes may be derivatized with various polymers (E.g., U.S. Patent No. 5,013,556). A description of possible solid dosage forms for the therapeutic is given by Marshall, K. In: Modern Pharmaceutics Edited by G.S. Banker and CT. Rhodes Chapter 10, 1979, herein incorporated by reference. In general, the formulation will include the protein (or analog or derivative) , and inert ingredients which allow for protection against the stomach environment, and release of the biologically active material in the intestine. Also specifically contemplated are oral dosage forms of the above derivatized proteins. Protein may be chemically modified so that oral delivery of the derivative is efficacious. Generally, the chemical modification contemplated is the attachment of at least one moiety to the protein (or peptide) molecule itself, where said moiety permits (a) inhibition of proteolysis; and (b) uptake into the blood stream from the stomach or intestine. Also desired is the increase in overall stability of the protein and increase in circulation time in the body. Examples of such moieties include: Polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone and polyproline. Abuchowski and Davis, Soluble Polymer-Enzyme Adducts . In: "Enzymes as Drugs", Hocenberg and Roberts, eds . , Wiley-Interscience, New York, NY, (1981), pp 367-383; New ark, et al . , J. Appl . Biochem. A: 185-189 (1982) . Other polymers that could be used are poly-1, 3-dioxolane and poly-1, 3 , 6-tioxocane. For the protein (or derivative) the location of release may be the stomach, the small intestine (the duodenum, the jejunem, or the ileum) , or the large intestine. One skilled in the art has available formulations which will not dissolve in the stomach, yet will release the material in the duodenum or elsewhere in the intestine. Preferably, the release will avoid the deleterious effects of the stomach environment, either by protection of the protein (or derivative) or by release of the biologically active material beyond the stomach environment, such as in the intestine. To ensure full gastric resistance a coating impermeable to at least pH 5.0 is essential . Examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT) , hydroxypropylmethylcellulose phthalate (HPMCP) , HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP) , Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and Shellac. These coatings may be used as mixed films. A coating or mixture of coatings can also be used on tablets, which are not intended for protection against the stomach. This can include sugar coatings, or coatings which make the tablet easier to swallow. Capsules may consist of a hard shell (such as gelatin) for delivery of dry therapeutic i.e. powder; for liquid forms, a soft gelatin shell may be used. The shell material of cachets could be thick starch or other edible paper. For pills, lozenges, molded tablets or tablet triturates, moist massing techniques can be used. The therapeutic can be included in the formulation as fine multiparticulates in the form of granules or pellets of particle size about 1mm. The formulation of the material for capsule administration could also be as a powder, lightly compressed plugs or even as tablets . The therapeutic could be prepared by compression. Colorants and flavoring agents may all be included. For example, the protein (or derivative) may be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavoring agents. One may dilute or increase the volume of the therapeutic with an inert material . These diluents could include carbohydrates, especially mannitol, a-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch. Certain inorganic salts may be also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell. Disintegrants may be included in the formulation of the therapeutic into a solid dosage form. Materials used as disintegrates include but are not limited to starch including the commercial disintegrant based on starch, Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may all be used. Another form of the disintegrants are the insoluble cationic exchange resins. Powdered gums may be used as disintegrants and as binders and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants. Binders may be used to hold the therapeutic agent together to form a hard tablet and include materials from natural products such as acacia, tragacanth, starch and gelatin. Others include methyl cellulose (MC) , ethyl cellulose (EC) and carboxymethyl cellulose (CMC) . Polyvinyl pyrrolidone (PVP) and hydroxypropylmethy1 cellulose (HPMC) could both be used in alcoholic solutions to granulate the therapeutic. An antifrictional agent may be included in the formulation of the therapeutic to prevent sticking during the formulation process . Lubricants may be used as a layer between the therapeutic and the die wall, and these can include but are not limited to; stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE) , liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000. Glidants that might improve the flow properties of the drug during formulation and to aid rearrangement during compression might be added. The glidants may include starch, talc, pyrogenic silica and hydrated silicoaluminate. To aid dissolution of the therapeutic into the aqueous environment a surfactant might be added as a wetting agent. Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents might be used and could include benzalkonium chloride or benzethomium chloride. The list of potential nonionic detergents that could be included in the formulation as surfactants are lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants could be present in the formulation of the protein or derivative either alone or as a mixture in different ratios. Additives which potentially enhance uptake of the protein (or derivative) are for instance the fatty acids oleic acid, linoleic acid and linolenic acid. Controlled release formulation may be desirable. The drug could be incorporated into an inert matrix which permits release by either diffusion or leaching mechanisms i.e. gums. Slowly degenerating matrices may also be incorporated into the formulation. Another form of a controlled release of this therapeutic is by a method based on the Oros therapeutic system (Alza Corp.), i.e. the drug is enclosed in a semipermeable membrane which allows water to enter and push drug out through a single small opening due to osmotic effects. Some entric coatings also have a delayed release effect. Other coatings may be used for the formulation. These include a variety of sugars which could be applied in a coating pan. The therapeutic agent could also be given in a film coated tablet and the materials used in this instance are divided into 2 groups. The first are the nonenteric materials and include methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, methylhydroxy-ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl-methyl cellulose, sodium carboxy-methyl cellulose, providone and the polyethylene glycols. The second group consists of the enteric materials that are commonly esters of phthalic acid. A mix of materials might be used to provide the optimum film coating. Film coating may be carried out in a pan coater or in a fluidized bed or by compression coating. Also contemplated herein is pulmonary delivery of the present protein, or derivative thereof. The protein (derivative) is delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream. (Other reports of this include Adjei et al . , Pharmaceutical Research 7: 565-569 (1990); Adjei et al . , International Journal of Pharmaceutics £2: 135-144 (1990) (leuprolide acetate); Braquet et al . , Journal of Cardiovascular Pharmacology 13 (suppl . 5): s.143-146 (1989) (endothelin-1) ;Hubbard et al . , Annals of Internal Medicine 2: 206-212 (1989) (al-antitrypsin) ; Smith et al., J. Clin. Invest.84.: 1145-1146 (1989) (a-1-proteinase) ; Oswein et al., "Aerosolization of Proteins", Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, 1990 (recombinant human growth hormone); Debs et al . , The Journal of Immunology 140: 3482-3488 (1988) (interferon-g and tumor necrosis factor alpha) and Platz et al . , U.S. Patent No. 5,284,656 (granulocyte colony stimulating factor) . Contemplated for use in the practice of this invention are a wide range of mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art. Some specific examples of commercially available devices suitable for the practice of this invention are the Ultravent nebulizer, manufactured by Mallinckrodt, Inc., St. Louis, Missouri; the Acorn II nebulizer, manufactured by Marquest Medical Products, Englewood, Colorado; the Ventolin metered dose inhaler, manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and the Spinhaler powder inhaler, manufactured by Fisons Corp., Bedford, Massachusetts. All such devices require the use of formulations suitable for the dispensing of protein (or analog or derivative) . Typically, each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material, in addition to diluents, adjuvants and/or carriers useful in therapy. The protein (or derivative) should most advantageously be prepared in particulate form with an average particle size of less than 10 m m (or microns) , most preferably 0.5 to 5 m m, for most effective delivery to the distal lung. Carriers include carbohydrates such as trehalose, mannitol, xylitol, sucrose, lactose, and sorbitol. Other ingredients for use in formulations may include DPPC, DOPE, DSPC and DOPC . Natural or synthetic surfactants may be used. Polyethylene glycol may be used (even apart from its use in derivatizing the protein or analog) . Dextrans , such as cyclodextran, may be used. Bile salts and other related enhancers may be used. Cellulose and cellulose derivatives may be used. Amino acids may be used, such as use in a buffer formulation. Also, the use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers is contemplated. Formulations suitable for use with a nebulizer, either jet or ultrasonic, will typically comprise protein (or derivative) dissolved in water at a concentration of about 0.1 to 25 mg of biologically active protein per mL of solution. The formulation may also include a buffer and a simple sugar (e.g., for protein stabilization and regulation of osmotic pressure) . The nebulizer formulation may also contain a surfactant, to reduce or prevent surface induced aggregation of the protein caused by atomization of the solution in forming the aerosol. Formulations for use with a etered-dose inhaler device will generally comprise a finely divided powder containing the protein (or derivative) suspended in a propellant with the aid of a surfactant. The propellant may be any conventional material employed for this purpose, such as a chlorofluorocarbon, a hydrochlorofluorocarbon, a hydrofluorocarbon, or a hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol , and 1, 1, 1, 2-tetrafluoroethane, or combinations thereof. Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid may also be useful as a surfactant . Formulations for dispensing from a powder inhaler device will comprise a finely divided dry powder containing protein (or derivative) and may also include a bulking agent, such as lactose, sorbitol, sucrose, mannitol, trehalose, or xylitol in amounts which facilitate dispersal of the powder from the device, e.g. , 50 to 90% by weight of the formulation. Nasal delivery of the protein (or analog or derivative) is also contemplated. Nasal delivery allows the passage of the protein to the blood stream directly after administering the therapeutic product to the nose, without the necessity for deposition of the product in the lung. Formulations for nasal delivery include those with dextran or cyclodextran. Delivery via transport across other mucus membranes is also contemplated. Dosages One skilled in the art will be able to ascertain effective dosages by administration and observing the desired therapeutic effect. The present invention provides that the dosages of OB protein alone required for a given effect will be reduced when a composition increasing the sensitivity of the OB receptor, e.g., increasing the affinity of the OB receptor to it's ligand (OB protein) or "upregulating" OB protein receptor, is also provided. Preferably, the formulation of the molecule will be such that between about .10 μg/kg/day and 100 mg/kg/day will yield the desired therapeutic effect. The effective dosages may be determined using diagnostic tools over time. For example, a diagnostic for measuring the amount of OB protein in the blood (or plasma or serum) may first be used to determine endogenous levels of OB protein. Such diagnostic tool may be in the form of an antibody assay, such as an antibody sandwich assay. The amount of endogenous OB protein is quantified initially, and a baseline is determined. The therapeutic dosages are determined as the quantification of endogenous and exogenous OB protein (that is, protein, analog or derivative found within the body, either self-produced or administered) is continued over the course of therapy. The dosages may therefore vary over the course of therapy, with a relatively high dosage being used initially, until therapeutic benefit is seen, and lower dosages used to maintain the therapeutic benefits. Ideally, in situations where solely an increase in lean body mass is desired, the dosage will be insufficient to result in weight loss. Thus, during an initial course of therapy of an obese person, dosages may be administered whereby weight loss and concomitant fat tissue decrease/lean mass increase is achieved. Once sufficient weight loss is achieved, a dosage sufficient to prevent re-gaining weight, yet sufficient to maintain desired lean mass increase (or, prevention of lean mass depletion) may be administered. These dosages can be determined empirically, as the effects of OB protein are reversible. E.g. , Ca pfield et al . , Science 269 : 546-549 (1995) at 547. Thus, if a dosage resulting in weight loss is observed when weight loss is not desired, one would administer a lower dose in order to achieve the desired increase in lean tissue mass, yet maintain the desired weight. For increasing an individual's sensitivity to insulin, similar dosage considerations may be taken into account. Lean mass increase without weight loss may be achieved sufficient to decrease the amount of insulin (or, potentially, amylin or other potential diabetes treating drugs) an individual would be administered for the treatment of diabetes . For increasing overall strength, there may be similar dosage considerations. Lean mass increase with concomitant increase in overall strength may be achieved with doses insufficient to result in weight loss. Other benefits, such as an increase in red blood cells (and oxygenation in the blood) and a decrease in bone resorption or osteoporosis may also be achieved in the absence of weight loss. Combinations The present methods may be used in conjunction with other medicaments, such as those useful for the treatment of diabetes (e.g., insulin, possibly thiazolidinediones, amylin or antagonists thereof) , cholesterol and blood pressure lowering medicaments (such as those which reduce blood lipid levels or other cardiovascular medicaments), and activity increasing medicaments (e.g., amphetamines). Appetite suppressants may also be used (such as those affecting the levels of serotonin or neuropeptide Y) . Such administration may be simultaneous or may be in seriatim. In addition, the present methods may be used in conjunction with surgical procedures, such as cosmetic surgeries designed to alter the overall appearance of a body (e.g., liposuction or laser surgeries designed to reduce body mass, or implant surgeries designed to increase the appearance of body mass). The health benefits of cardiac surgeries, such as bypass surgeries or other surgeries designed to relieve a deleterious condition caused by blockage of blood vessels by fatty deposits, such as arterial plaque, may be increased with concomitant use of the present compositions and methods . Methods to eliminate gall stones, such as ultrasonic or laser methods, may also be used either prior to, during or after a course of the present therapeutic methods. Furthermore, the present methods may be used as an adjunct to surgeries or therapies for broken bones, damaged muscle, or other therapies which would be improved by an increase in lean tissue mass. Therefore, the present invention provides a method for increasing, in an individual, the sensitivity to OB protein or analog or derivative thereof, comprised of administering an effective amount of a composition which increases the availability, affinity, or sensitivity of functional OB protein receptor in said individual. Optionally, said method also involves the administration, either simultaneously or m . serriatim of an OB protein, analog or derivative thereof. Said composition which increases the availability or sensitivity of functional OB protein receptor may be selected from among thiazolidinedione compositions: 2 , 4-thiazolidinediones; opt. substituted thiazolidinediones; 5- [4- [2- (5-methyl-2-phenyl-4- oxazolyl) -2-hydroxyethoxy] enzyl] -2 , 4-thiazolidinedione (AD-5070); clofibrate; ciglitazone; englitazone; pioglitazone; BRL 49653; troglitazone; M16209; oxazolidinediones; as well as derivatives, analogs, tautomers , enantiomers, diastereomers, epimers, salts, solvates, esters, prodrugs and metabolites of thiazolidinediones or the compounds above. The OB protein, analog or derivative thereof may be selected from among: (a) the amino acid sequence 1-146 as set forth in SEQ. ID. NO. 3 (below) or SEQ ID. NO. 6 (below) , (b) the amino acid sequence set 1-146 as forth in SEQ. ID. NO. 6 (below) having a lysine residue at position 35 and an isoleucine residue at position 74; (c) the amino acid sequence of subpart (b) having a different amino acid substituted in one or more of the following positions (using the numbering according to SEQ. ID. NO. 6, and retaining the same numbering even in the absence of a glutammyl residue at position 28): 4, 8, 32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145; (d) the amino acid sequence of subpartε (a) , (b) or (c) optionally lacking a glutammyl residue at position 28; (e) the amino acid sequence of subparts (a) , (b) , (c) , or (d) having a methionyl reεidue at the N terminus . (f) a truncated OB protein analog selected from among: (using the numbering of SEQ. ID. NO. 6 having a lysine residue at position 35 and an isoleucine residue at position 74) : (i) amino acids 98-146 (ii) amino acids 1-32 (iii) amino acids 40-116 (iv) amino acids 1-99 and 112-146 (v) amino acids 1-99 and 112-146 having one or more of amino acids 100-111 sequentially placed between amino acids 99 and 112; and, (vi) the truncated OB analog of subpart (i) having one or more of amino acids 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145 substituted with another amino acid; (vii) the truncated analog of subpart (ii) having one or more of amino acids 4, 8 and 32 substituted with another amino acid; (viii) the truncated analog of subpart (iii) having one or more of amino acids 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111 and 112 replaced with another amino acid; (vix) the truncated analog of subpart (iv) having one or more of amino acids 4, 8, 32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 112, 118, 136, 138, 142, and 145 replaced with another amino acid; (x) the truncated analog of subpart (v) having one or more of amino acids 4, 8,32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145 replaced with another amino acid; (xi) the truncated analog of any of subparts (i)-(x) having an N-terminal methionyl reεidue; and (g) the OB protein or analog derivative of any of subparts (a) through (f) comprised of a chemical moiety connected to the protein moiety; (h) a derivative of subpart (g) wherein said chemical moiety is a water soluble polymer moiety; (i) a derivative of subpart (h) wherein said water soluble polymer moiety is polyethylene glycol; (j) A derivative of subpart (h) wherein said water soluble polymer moiety is a polyamino acid moiety; (k) a derivative of subpart (h) wherein said water soluble polymer moiety is attached at solely the N-terminus of said protein moiety (1) an OB protein, analog or derivative of any of subparts (a) through (k) in a pharmaceutically acceptable carrier. The following example is offered to more fully illustrate the invention, but is not to be construed as limiting the scope thereof. Example 1 is a prophetic example of human use which demonstrates that thiazolidinedione compositions increase the affinity or availability of OB protein receptors in an individual and those individuals have an increased sensitivity to said OB protein. Materials and Methods follow. EXAMPLE 1 An obese human patient (BMI > 27) desires weight loss. The patient is administered an amount of a thiazolidinedione composition effective to increase the availability of OB protein receptor for one week. The patient is then dosed with an amount of OB protein, or derivative or analog thereof, sufficient to result in decrease in weight. Levels of circulating OB protein or analog or derivative may be monitored using a diagnostic kit, such as an antibody assay against the OB protein (or other antigenic source if applicable) . The patient loses weight, and obtains a desired body weight and/or fat mass . OB protein or analog or derivative thereof optionally in combination with a thiazolidinedione composition is thereafter chronically administered for a desired period of time to maintain the desired weight and/or body fat level. MATERIALS AND METHODS Administration of Protein or Vehicle. Protein: Sequence ID Nos. 1, 2 and 3 set forth murine recombinant OB DNA and protein (figure 1) , and Sequence ID Nos. 4, 5 and 6 set forth an analog recombinant human OB DNA and protein (figure 2) . Recombinant human OB protein as in SEQ. ID. NO. 6 having a lysine residue at position 35 and an isoleucine residue at position 74 was used in EXAMPLE 1. As indicated above, the below murine and human analog recombinant proteins are illustrative of the OB protein which may be used in the present methods of treatment and manufacture of a medicament . Other OB proteins or analogs or derivatives thereof may be used. Herein, the first amino acid of the amino acid sequence for recombinant protein is referred to as +1, and is valine, and the amino acid at position -1 is methionine. The C-terminal amino acid is number 146 (cysteine) . METHODS FOR PRODUCTION The below methods for production have been used to produce biologically active recombinant methionyl murine or human analog OB protein. Similar methods may be used to prepare biologically active recombinant methionyl human OB protein. Expression Vector and Host Strain The plasmid expression vector used is PCFM1656, ATCC Accession No. 69576. The above DNA was ligated into the expression vector pCFM1656 linearized with Xbal and BamHI and transformed into the E. coli host εtrain, FM5. E. coli FM5 cells were derived at Amgen Inc., Thousand Oaks, CA from E. coli K-12 strain (Bachmann, et al . , Bacteriol. Rev. 40 . : 116-167 (1976)) and contain the integrated lambda phage repressor gene, cl857 (Susεman et al . , CR. Acad. Sci. 254 : 1517-1579 (1962) ) . Vector production, cell transformation, and colony selection were performed by standard methods. E.g. , Sambrook, et al . , Molecular Cloning: A Laboratory Manual, 2d Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Host cells were grown in LB media . Fermentation Process A three-phase fermentation protocol known as a fed-batch process was used. Media compositions are set forth below. Batch: A nitrogen and phosphate source were sterilized (by raising to 122 °C for 35 minutes, 18-20 psi) in the fermentation vessel (Biolafitte, 12 liter capacity) . Upon cooling, carbon, magnesium, vitamin, and trace metal sources were added aseptically. An overnight culture of the above recombinant murine protein-producing bacteria (16 hours or more) of 500 mL (grown in LB broth) was added to the fermentor. Feed I: Upon reaching between 4.0-6.0 OD600' cultures were fed with Feed I. The glucoεe was fed at a limiting rate in order to control the growth rate (m) . An automated system (called the Distributive Control System) was instructed to control the growth rate to 0.15 generations per hour. Feed II: When the OD600 had reached 30, culture temperature were slowly increased to 42°C and the feed changed to Feed II, below. The fermentation was allowed to continue for 10 hours with sampling every 2 hours. After 10 hours, the contents of the fermentor was chilled to below 20°C and harvested by centrifugation . Media Composition: Batch: 10 g/L Yeast extract 5.25 g/L (NH4)2S04 3.5 g/L K2HPO4 4.0 g/L KH2PO4 5.0 g/L Glucose 1.0 g/L MgS04 • 7H20 2.0 mL/L Vitamin Solution 2.0 mL/L Trace Metal Solution 1.0 mL/L P2000 Antifoam Feed I: 50 g/L Bacto-tryptone 50 g/L Yeast extract 450 g/L Glucose 8.75 g/L MgS04 • 7H20 10 mL/L Vitamin Solution 10 mL/L Trace Metal Solution Feed II: 200 g/L Bacto-tryptone 100 g/L Yeast extract 110 g/L Glucose Vitamin Solution (Batch and Feed I) : 0.5 g Biotin, 0.4 g Folic acid, and 4.2 g riboflavin, was dissolved in 450 is H2O and 3 mis 10 N NaOH, and brought to 500 mLs in H2O. 14 g pyridoxine-HCl and 61 g niacin was dissolved 150 ml H2O and 50 ml 10 N NaOH, and brought to 250 ml in H2O. 54 g pantothenic acid was dissolved in 200 mL H2O, and brought to 250 mL. The three solutions were combined and brought to 10 liters total volume. Trace Metal Solution (Batch and Feed I) : Ferric Chloride (FeCl3 • 6H2O) : 27 g/L Zinc Chloride (ZnCl2 ■ 4H20) : 2 g/L Cobalt Chloride (C0CI2 ■ 6H2O) : 2 g/L Sodium Molybdate (NaMoθ4 • 2H20) : 2 g/L Calcium Chloride (CaCl2 • 2H20) : 1 g/L Cupric Sulfate (CUSO4 • 5H20) : 1.9 g/L Boric Acid (H3BO3): 0.5 g/L Manganese Chloride (MnCl2 • H20) : 1.6 g/L Sodium Citrate dihydrate: 73.5 g/L Purification Process for Murine OB Protein Purification was accomplished by the following steps (unlesε otherwise noted, the following steps were performed at 4°C) : 1. Cell paste. E. coli cell paste was suspended in 5 times volume of 7 mM of EDTA, pH 7.0. The cells in the EDTA were further broken by two passes through a microfluidizer . The broken cells were centrifuged at 4.2 K rpm for 1 hour in a Beckman J6-B centrifuge with a JS-4.2 rotor. 2. Inclusion body wash #1. The supernatant from above was removed, and the pellet was resuspended with 5 times volume of 7 mM EDTA, pH 7.0, and homogenized. This mixture was centrifuged as in step 1. 3. Inclusion body wash #2. The supernatant from above was removed, and the pellet was resuspended in ten times volume of 20 mM tris, pH 8.5 , 10 mM DTT, and 1% deoxycholate, and homogenized. This mixture waε centrifuged as in step 1. 4. Inclusion body wash #3. The supernatant from above was removed and the pellet was resuspended in ten times volume of distilled water, and homogenized. This mixture was centrifuged as in step 1. 5. Refolding. The pellet was refolded with 15 volumes of 10 mM HEPES, pH 8.5 , 1% sodium sarcosine (N-lauroyl sarcosine), at room temperature. After 60 minutes, the solution was made to be 60 mM copper sulfate, and then stirred overnight. 6. Removal of sarcosine. The refolding mixture was diluted with 5 volumes of 10 mM tris buffer, pH 7.5, and centrifuged as in step 1. The supernatant was collected, and mixed with agitation for one hour with Dowex® 1-X4 resin (Dow Chemical Co., Midland MI), 20-50 mesh, chloride form, at 0.066% total volume of diluted refolding mix. See WO 89/10932 at page 26 for more information on Dowex®. This mixture was poured into a column and the eluant collected. Removal of sarcosine was aεcertained by reverse phase HPLC 7. Acid precipitation. The eluant from the previous step was collected, and pH adjusted to pH 5.5, and incubated for 30 minutes at room temperature. This mixture was centrifuged as in step 1. 8. Cation exchange chromatography. The pH of the supernatant from the previous step was adjusted to pH 4.2, and loaded on CM Sepharose Fast Flow (at 7% volume) . 20 column volumes of salt gradient were done at 20 mM NaOAC, pH 4.2, O to l.O M NaCl . 9. Hydrophobic interaction chromatography. The CM Sepharose pool of peak fractions (ascertained from ultraviolet absorbance) from the above step was made to be 0.2 M ammonium sulfate. A 20 column volume reverse salt gradient was done at 5 mM NaOAC, pH 4.2, with .4 M to 0 M ammonium sulfate. This material was concentrated and diafiltered into PBS. Fermentation of recombinant human OB protein analo : Fermentation of the above host cells to produce recombinant human OB protein analog (SEQ. ID. NO. 6) can be- accomplished using the conditions and compositions as described above for recombinant murine material . Purification of the recombinant human OB protein analog: Recombinant human protein analog may be purified using methods similar to thoεe used for purification of recombinant murine protein, aε described above. For preparation of recombinant human OB protein analog, step 8 should be performed by adjusting the pH of the supernatant from step 7 to pH 5.0, and loading this onto a CM Sepharose fast flow column. The 20 column volume salt gradient should be performed at 20 mM NaOAC, pH 5.5, 0M to 0.5 M NaCl . Step 9 should be performed by diluting the CM Sepharose pool four fold with water, and adjusting the pH to 7.5. This mixture should be made to 0.7 M ammonium sulfate. Twenty column volume reverse salt gradient should be done at 5 mM NaOAC, pH 5.5 , 0.2 M to 0M ammonium sulfate. Otherwise, the above steps are identical. The recombinant human OB protein of SEQ. ID.NO.6 having lysine 35 and isoleucine 74 was formulated in a buffer containing 10 mM histidine, 4.3% arginine, at pH 6.0. While the present invention has been described in terms of preferred embodiments, it is understood that variations and modifications will occur to those skilled in the art. Therefore, it is intended that the appended claims cover all such equivalent variations which come within the scope of the invention as claimed. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: AMGEN INC. (ii) TITLE OF INVENTION: METHODS OF INCREASING SENSITIVITY OF AN INDIVIDUAL TO OB PROTEIN BY UPREGULATING OB PROTEIN RECEPTOR (iii) NUMBER OF SEQUENCES: 6 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: AMGEN INC. (B) STREET: 1840 DEHAVILLAND DRIVE (C) CITY: THOUSAND OAKS (D) STATE: CA (E) COUNTRY: USA (F) ZIP: 91320-1789 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.30 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: 08/705,981 (B) FILING DATE: 30 August 1996 (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: KNIGHT, MATTHEW W (B) REGISTRATION NUMBER: 36,846 (C) REFERENCE/DOCKET NUMBER: A-421 (2) INFORMATION FOR SEQ ID NO : 1 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 491 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 1 : TCTAGATTTG AGTTTTAACT TTTAGAAGGA GGAATAACAT ATGGTACCGA TCCAGAAAGT 60 TCAGGACGAC ACCAAAACCT TAATTAAAAC GATCGTTACG CGTATCAACG ACATCAGTCA 120 CACCCAGTCG GTCTCCGCTA AACAGCGTGT TACCGGTCTG GACTTCATCC CGGGTCTGCA 180 CCCGATCCTA AGCTTGTCCA AAATGGACCA GACCCTGGCT GTATACCAGC AGGTGTTAAC 240 CTCCCTGCCG TCCCAGAACG TTCTTCAGAT CGCTAACGAC CTCGAGAACC TTCGCGACCT 300 GCTGCACCTG CTGGCATTCT CCAAATCCTG CTCCCTGCCG CAGACCTCAG GTCTTCAGAA 360 ACCGGAATCC CTGGACGGGG TCCTGGAAGC ATCCCTGTAC AGCACCGAAG TTGTTGCTCT 420 GTCCCGTCTG CAGGGTTCCC TTCAGGACAT CCTTCAGCAG CTGGACGTTT CTCCGGAATG 480 TTAATGGATC C 491 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 491 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE- TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: AGATCTAAAC TCAAAATTGA AAATCTTCCT CCTTATTGTA TACCATGGCT AGGTCTTTCA 60 AGTCCTGCTG TGGTTTTGGA ATTAATTTTG CTAGCAATGC GCATAGTTGC TGTAGTCAGT 120 GTGGGTCAGC CAGAGGCGAT TTGTCGCACA ATGGCCAGAC CTGAAGTAGG GCCCAGACGT 180 GGGCTAGGAT TCGAACAGGT TTTACCTGGT CTGGGACCGA CATATGGTCG TCCACAATTG 240 GAGGGACGGC AGGGTCTTGC AAGAAGTCTA GCGATTGCTG GAGCTCTTGG AAGCGCTGGA 300 CGACGTGGAC GACCGTAAGA GGTTTAGGAC GAGGGACGGC GTCTGGAGTC CAGAAGTCTT 360 TGGCCTTAGG GACCTGCCCC AGGACCTTCG TAGGGACATG TCGTGGCTTC AACAACGAGA 420 CAGGGCAGAC GTCCCAAGGG AAGTCCTGTA GGAAGTCGTC GACCTGCAAA GAGGCCTTAC 480 AATTACCTAG G 491 ) INFORMATION FOR SEQ ID NO : 3 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 147 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( ix ) FEATURE : (A) NAME/KEY: Protein (B) LOCATION: 1..147 (D) OTHER INFORMATION: /note= "NOTE: METHIONYL RESIDUE STARTS AT THE -1 POSITION." (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 3 : Met Val Pro lie Gin Lys Val Gin Asp Asp Thr Lys Thr Leu lie Lys 1 5 10 15 Thr lie Val Thr Arg lie Asn Asp lie Ser His Thr Gin Ser Val Ser 20 25 30 Ala Lys Gin Arg Val Thr Gly Leu Asp Phe lie Pro Gly Leu His Pro 35 40 45 lie Leu Ser Leu Ser Lys Met Asp Gin Thr Leu Ala Val Tyr Gin Gin 50 55 60 Val Leu Thr Ser Leu Pro Ser Gin Asn Val Leu Gin lie Ala Asn Asp 65 70 75 80 Leu Glu Asn Leu Arg Asp Leu Leu His Leu Leu Ala Phe Ser Lys Ser 85 90 95 Cys Ser Leu Pro Gin Thr Ser Gly Leu Gin Lys Pro Glu Ser Leu Asp 100 105 110 Gly Val Leu Glu Ala Ser Leu Tyr Ser Thr Glu Val Val Ala Leu Ser 115 120 125 Arg Leu Gin Gly Ser Leu Gin Asp lie Leu Gin Gin Leu Asp Val Ser 130 135 140 Pro Glu Cys 145 (2) INFORMATION FOR SEQ ID NO : 4 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 454 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: CATATGGTAC CGATCCAGAA AGTTCAGGAC GACACCAAAA CCTTAATTAA AACGATCGTT 60 ACGCGTATCA ACGACATCAG TCACACCCAG TCGGTGAGCT CTAAACAGCG TGTTACAGGC 120 CTGGACTTCA TCCCGGGTCT GCACCCGATC CTGACCTTGT CCAAAATGGA CCAGACCCTG 180 GCTGTATACC AGCAGATCTT AACCTCCATG CCGTCCCGTA ACGTTCTTCA GATCTCTAAC 240 GACCTCGAGA ACCTTCGCGA CCTGCTGCAC GTGCTGGCAT TCTCCAAATC CTGCCACCTG 300 CCATGGGCTT CAGGTCTTGA GACTCTGGAC TCTCTGGGCG GGGTCCTGGA AGCATCCGGT 360 TACAGCACCG AAGTTGTTGC TCTGTCCCGT CTGCAGGGTT CCCTTCAGGA CATGCTTTGG 420 CAGCTGGACC TGTCTCCGGG TTGTTAATGG ATCC 454 (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 454 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 5 : GTATACCATG GCTAGGTCTT TCAAGTCCTG CTGTGGTTTT GGAATTAATT TTGCTAGCAA 60 TGCGCATAGT TGCTGTAGTC AGTGTGGGTC AGCCACTCGA GATTTGTCGC ACAATGTCCG 120 GACCTGAAGT AGGGCCCAGA CGTGGGCTAG GACTGGAACA GGTTTTACCT GGTCTGGGAC 180 CGACATATGG TCGTCTAGAA TTGGAGGTAC GGCAGGGCAT TGCAAGAAGT CTAGAGATTG 240 CTGGAGCTCT TGGAAGCGCT GGACGACGTG CACGACCGTA AGAGGTTTAG GACGGTGGAC 300 GGTACCCGAA GTCCAGAACT CTGAGACCTG AGAGACCCGC CCCAGGACCT TCGTAGGCCA 360 ATGTCGTGGC TTCAACAACG AGACAGGGCA GACGTCCCAA GGGAAGTCCT GTACGAAACC 420 GTCGACCTGG ACAGAGGCCC AACAATTACC TAGG 454 (2) INFORMATION FOR SEQ ID NO : 6 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 147 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (ix) FEATURE: (A) NAME/KEY: Protein (B) LOCATION: 1..147 (D) OTHER INFORMATION: /note= "NOTE: METHIONYL RESIDUE STARTS AT THE -1 POSITION." (xi) SEQUENCE -DESCRIPTION: SEQ ID NO: 6: Met Val Pro He Gin Lys Val Gin Asp Asp Thr Lys Thr Leu He Lys 1 5 10 15 Thr He Val Thr Arg He Asn Asp He Ser His Thr Gin Ser Val Ser 20 25 30 Ser Lys Gin Arg Val Thr Gly Leu Asp Phe He Pro Gly Leu His Pro 35 40 45 He Leu Thr Leu Ser Lys Met Asp Gin Thr Leu Ala Val Tyr Gin Gin 50 55 60 He Leu Thr Ser Met Pro Ser Arg Asn Val Leu Gin He Ser Asn Asp 65 70 75 80 Leu Glu Asn Leu Arg Asp Leu Leu His Val Leu Ala Phe Ser Lys Ser 85 90 95 Cys His Leu Pro Trp Ala Ser Gly Leu Glu Thr Leu Asp Ser Leu Gly 100 105 110 Gly Val Leu Glu Ala Ser Gly Tyr Ser Thr Glu Val Val Ala Leu Ser 115 120 125 Arg Leu Gin Gly Ser Leu Gin Asp Met Leu Trp Gin Leu Asp Leu Ser 130 135 140 Pro Gly Cys 145

Claims: CLAIMS 1. A method for increasing, in an individual, the sensitivity to an OB protein or analog or derivative thereof, by administration of a composition which increases the affinity or availability of functional OB protein receptor within said individual, said method optionally in combination with administration of said OB protein or analog or derivative thereof. 2. A method of claim 1 wherein said composition which increases the affinity or availability of functional OB protein receptor is a thiazolidinedione composition. 3. A method of claim 1 or 2 wherein said OB protein or analog or derivative thereof is selected from among: (a) the amino acid sequence 1-146 as set forth in SEQ. ID. NO. 3 or SEQ ID. NO. 6 ; (b) the amino acid sequence 1-146 as set forth in SEQ. ID. NO. 6 having a lysine residue at position 35 and an isoleucine residue at position 74; (c) the amino acid sequence of subpart (b) having a different amino acid substituted in one or more of the following positions (using the numbering according to SEQ. ID. NO. 4): 4, 8, 32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145; (d) the amino acid sequence of subparts (a) , (b) or (c) optionally lacking a glutaminyl residue at position 28; (e) the amino acid sequence of subparts (a) , (b) , (c) , or (d) having a methionyl residue at the N terminus . (f) a truncated OB protein analog selected from among: (using the numbering of SEQ. ID. NO. 6 having a lysine residue at position 35, and an isoleucine residue at position 74): (i) amino acids 98-146 (ii) amino acids 1-32 (iii) amino acids 40-116 (iv) amino acids 1-99 and 112-146 (v) amino acids 1-99 and 112-146 having one or more of amino acids 100-111 sequentially placed between amino acids 99 and 112; and, (vi) the truncated OB analog of subpart (f) (i) having one or more of amino acids 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145 substituted with another amino acid; (vii) the truncated analog of subpart (f) (ii) having one or more of amino acids 4, 8 and 32 substituted with another amino acid; (viii) the truncated analog of subpart (f) (iii) having one or more of amino acids 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111 and 112 replaced with another amino acid; (vix) the truncated analog of subpart (f) (iv) having one or more of amino acids 4, 8, 32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 112, 118, 136, 138, 142, and 145 replaced with another amino acid; (x) the truncated analog of subpart (f) (v) having one or more of amino acids 4, 8,32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145 replaced with another amino acid; (xi) the truncated analog of any of subparts (f) (i)-(x) having an N-terminal methionyl residue; and (g) the OB protein or analog derivative of any of subparts (a) through (f) comprised of a chemical moiety connected to the protein moiety; (h) a derivative of subpart (g) wherein said chemical moiety is a water soluble polymer moiety; (i) a derivative of subpart (h) wherein said water soluble polymer moiety is polyethylene glycol; (j) A derivative of subpart (h) wherein said water soluble polymer moiety is a polyamino acid moiety; (k) a derivative of subpart (h) wherein said water soluble polymer moiety is attached at solely the N-terminus of said protein moiety; (1) an OB protein, analog or derivative of any of subparts (a) through (k) in a pharmaceutically acceptable carrier . 4. A method of claim 1, 2, or 3 wherein said administration is for the treatment for a condition selected from among excess weight, diabetes, high blood lipid level, artherial sclerosis, artherial plaque, the reduction or prevention of gall stone formation, insufficient lean tissue mass, insufficient sensitivity to insulin, and stroke.

Inventors: PELLEYMOUNTER MARY ANN, PELLEYMOUNTER, MARY, ANN
Applicants: AMGEN INC, AMGEN INC.
Application year: 1997
Main classification: A61K31
IPC codes: A61K31, A61K38, A61P1, A61P3, A61P9, C07K14