CETP INHIBITOR POLYPEPTIDE, ANTIBODIES AGAINST THE SYNTHETIC POLYPEPTIDE AND PROPHYLACTIC AND THERAPEUTIC ANTI-ATHEROSCLEROSIS TREATMENTS

Abstract: A polypeptide and analogues thereof inhibit cholesteryl ester transfer protein (CETP). An anti-atherosclerosis composition comprises an anti-atherosclerosis effective amount of the polypeptide and a pharmaceutically-acceptable carrier. An anti-atherosclerosis kit comprises in separate sterile containers at least one unit of the composition containing the polypeptide, one syringe and one needle. An antibody has specificity for the polypeptide of the invention, the baboon CETP 4kD polypeptide inhibitor, the 1-36 amino acid N-terminal fragment of apo C-I, modified apo A-I (MW:31kD) or modified apo E (MW:41kD). A method of preventing atherosclerosis in a mammal being predisposed to that condition comprises administering to the mammal a prophylactically effective amount of the polypeptide of the invention, and a method of treating a mammal afflicted with atherosclerosis comprises the administration of a therapeutically effective amount of the polypeptide disclosed herein.

Description: CETP INHIBITOR POLYPEPTIDE, ANTIBODIES AGAINST THE SYNTHETIC POLYPEPTIDE AND PROPHYLACTIC AND THERAPEUTIC ANTI-ATHEROSCLEROSIS TREATMENTS BACKGROUND OF THE INVENTION The work leading to the present invention was partially supported by National Heart, Lung and Blood Institute Grants Nos. HL28972 and HL41256, and Contract No. HV53030. The government may hold rights in the present patent. Field of the Invention This invention relates to an endogenous baboon plasma cholesteryl esters transfer protein (CETP) inhibitor polypeptide. More specifically, this invention relates to the identification and characterization of the polypeptide and to novel synthetic peptides possessing inhibitory activity of CETP. The endogenous inhibitory peptide has a molecular weight of 4000, is present in plasma in the form of modified apo A-I and apo E having molecular weights of 31kD and 41kD, respectively, and has a common amino acid sequence with the N-terminal fragment of apo C-I. This invention also relates to an anti- atherosclerosis composition, a kit, and to antibodies raised against the N-teπninal amino acid sequence of the inhibitory polypeptide. The inhibitory peptide of the invention, fragments thereof and analogues thereof are useful for the prophylactic and therapeutic treatment of atherosclerosis. Description of the Background Atherosclerosis is one of the most widespread health problems in the United States today as are its attendant complications, particularly coronary heart disease. A number of risk factors have been associated with the development of premature atherosclerosis, primarily elevated plasma cholesterol levels. Due to the crucial role cholesterol appears to play in the occurrence of heart disease, a great deal of attention has been devoted to studying its synthesis, transport and metabolism in the human body. Of particular interest is the establishment of relationships between the levels of plasma lipoproteins or serum lipids and the risk of development of coronary heart disease. Both high density lipoproteins (HDL) and low density lipoproteins (LDL) carry cholesterol mainly in the form of cholestryl esters (CE). There are some indications, however, that while LDL cholesterol is a positive risk factor, HDL cholesterol is an even more important negative risk factor. Although the exact functions of these lipoproteins have not been completely established, HDL appears to serve for the removal of cholesterol from peripheral cells and its transport back to the liver, where a large proportion of the cholesterol excreted from the body is removed. LDL and HDL are believed to play key roles in the development of cardiovascular disease by overloading the lysosomes of the walls of arterial cells with metabolites which are generally hydrolyzed slowly, such as CE and triglycerides. These products are evacuated from the liver and intestine by plasma LDL. When the amount of lipids to be transported exceeds the transporting capacity of HDL to the liver for excretion, CE become deposited in the cells in certain critical areas, such as arterial walls. This overloading eventually results in impaired cell function, and if continued may produce cell death. A continuous overloading results in the accumulation of cellular debris and the formation of atherosclerotic plaque in the vessel wall. This, in turn, leads to the blockage of the affected artery and/or muscular spasm, events which may manifest themselves as coronary heart disease or strokes. Thus, the level of HDL in plasma has been negatively correlated with the probability of developing atherosclerosis in humans and experimental animals. Although the level of HDL has been shown to vary considerably among individuals, the means of regulation of such plasma level remains to be elucidated. CETP transfers CE from HDL to VLDL and LDL, and it has been suggested that it plays an important role in the regulation of plasma HDL levels. Some hyperalphaHpo-proteinemic patients were reported to have high levels of large HDL particles that were clearly separate from LDL. Plasma samples from these patients were shown to lack CETP activity (Koizami et al, Atherosclerosis 58:175-186(1986)). A homozygous subject with familial hyperalphaHpoproteinemia was found to have impaired transfer of CE from HDL to LDL (Yokoyama et al., Artery 14:43-51(1986)). A fraction of density d 1 1.21 g/ml from the subject's plasma evidenced substantial CETP activity with normal HDL. The HDL, however, proved to be a poor substrate for CETP. Certain animal sires and their progeny possess unusual lipoproteins patterns, e.g., Hpoproteins of a density intermediate to that of LDL and HDL, or large high density hpoproteins. These Hpoproteins have been designated HDL-; , and the animal phenotype as "high HDLi". Baboon strains possessing, for instance, patterns of either high or low HDLi are known. In most cases, HDL-j separates either as a distinct peak between LDL and HDL or as a shoulder to the HDL peak, and is induced by a high cholesterol, high lard (HCHF) diet. The proportion of HDLi diminishes when the baboons are fed a diet that is either enriched in polyunsaturated fat, with or without cholesterol. Occasionally, however, the amount of HDLi present in high HDL baboons fed the chow diet is low. In some baboon families, the level of plasma HDL was shown to increase when the animals are chaHenged with a HCHF diet. When fed a HCHF diet, the baboons also show higher plasma HDL. More generaUy, the accumulation of HDL in baboons as weU as in humans is associated with a slower transfer of CE from HDL to very low density Hpoproteins (VLDL) and LDL. Thus, baboons with high HDLi plasma levels are exceUent as animal models for the study of hyperalphaHpoproteinemia. In a previous study, some of the present inventors reported that a slower transfer of CE from HDL to VLDL and LDL was observed in high HDLi baboons. This was attributed to the presence of a CETP protein inhibitor associated with HDL and intermediate density Hpoprotein (EDL) particles (Kushwaha et al, JX. Lipid Res. 31:965-974(1990)). An accumulation of HDLi -** 11 tne ^-Φ 1 HDLi baboons fed a HCHF diet was reported along with a slower transfer of CE from HDL to LDL. A similar protein was found in human plasma by Son and Zilversmit (Son and Zilversmit, B.B.A. 795:473-480(1984)). The human protein has a molecular weight of 31,000 and suppresses the transfer of triacylglycerol and CE. Several other species including rat, pig and dog, have been reported to readily accumulate HDL ^ plasma. Kushwaha et al (1990), supra, reported that a homozygous subject with familial hyperalphaHpoproteinemia, has impaired CE transfer between HDL and LDL (Kushwaha et al, J.B. Biochem. 98:1499-1508(1985)). Separately, Yokoyama et al reported that a plasma fraction of ά 1 1.21 g/ml of the same subject evidenced substantial CE transfer activity when tested with normal HDL (Yokoyama et al, Artery 14(l):43-51(1986)). The HDL particles accumulated by this subject were substantiaUy larger in molecular size than ordinary HDL2. HDL is generaHy divided into subtractions based on their particle sizes and densities. These fractions include HDLi, HDL2 and HDL3. HDLi nas th e largest particles and is usuaHy not present in the plasma of normal humans or non-human primates. HDL2 and HDL3 are the normal components of human plasma. HDL2 is larger than HDL3 and differs between men and women. Many attempts have been made to interfere with the transport and transfer of cholesterol in mammaHans in order to alter its plasma levels. Among them are the foHowing. U.S. Patent No. 4,987,151 to Taboc discloses triterpene derivatives that inhibit acyl coenzyme A holesterol acyltransferase (ACAT) enzyme. The ACAT is a ceUular enzyme that is not present in plasma, and esterfies ceUular cholesterol to form CE. This enzyme is different from the CE transfer protein (CETP) present in plasma. The CETP does not form CE as does the ACAT enzyme. Instead, the CETP transfers CE amongst different plasma Hpoproteins. U.S. Patent No. 4,643,988 to Segrest et al discloses amphipathic peptides which are capable of substituting for apo A-I in HDL. Apo A-I is known to stimulate the lecithin cholesterol: acyl transferase (LCAT) enzyme, a plasma enzyme that forms CE in HDL. Plasma CETP, in contradistinction, transfers CE from HDL to VLDL and LDL. The function of the CETP enzyme is, therefore, different from that of the LCAT enzyme, as weU. The amino acid sequences of the Segrest et al peptides are, in addition, different from the sequences of the CETP inhibitor of this invention. SUMMARY OF THE INVENTION This invention relates to a substantiaHy pure polypeptide having activity inhibitory of CE transfer protein (CETP). This invention also relates to an anti-atherosclerosis composition, comprising an anti-atherosclerosis effective amount of the polypeptide described above; and a pharmaceuticaHy acceptable carrier. In addition, this invention relates to an anti-atherosclerosis kit, comprising in separate containers at least one unit of the composition described above; at least one syringe; and at least one needle. This invention also relates to an antibody having specificity for a polypeptide selected from the group consisting of the polypeptide described above; baboon CETP polypeptide inhibitor; 1-36 amino acid N-terminal fragment of apo C-I; modified apo A-I (MW:31kD); and modified apo E (MW:41kD). In a different aspect, this invention relates to a method of preventing atherosclerosis in a mammal being predisposed to that condition, comprising administering to the mammal a prophylacticaHy effective amount of the polypeptide described above. This invention also relates to a method of treating a mammal afflicted with atherosclerosis comprising administering to the mammal a therapeuticaUy effective amount of the polypeptide described above. Other objects, advantages and features of the present invention wiH become apparent to those skilled in the art from the foUowing discussion. DESCRIPTION OF THE PREFERRED EMBODIMENTS This invention arose from the desire by the inventors to provide a novel and unobvious approach to the prevention and treatment of atherosclerosis in humans. A schematic of what is known about the association of cholesterol with the different fractions of Hpoproteins in plasma and in the Hver is shown below. The metaboHc steps • leading to the accumulation (O) of HDLi, VLDL and LDL are shown above. HDL2+3 coHects cholesterol from extrahepatic ceUs, which is then esterified by LCAT to form cholesteryl esters (CE) and stored in the core of the particles. The HDL becomes larger in size (HDLi) and may pick up apo E to attain a particle form which is removed by LDL receptors (LDL-R) on Hver ceHs. The CE enriched HDLi may also donate CE to VLDL and LDL. This is mediated by CETP. Due to the presence of a CETP inhibitor, such as the one provided herein, CE transfer is slow (bar) and the reciprocal transfer of triglycerides (TG) does not take place (X). The triglyceride-poor HDL is thus not a suitable substrate for hepatic triglyceride Hpase (HTGL). Due to the presence of the CETP inhibitor, in plasma, VLDL and LDL are thus not available to the Hver. As a consequence of this, the Hver then increases the expression of messages for the increased production of LDL receptor and 3 hydroxy, methyl, glutaryl-coenzyme A (HMG-COA) synthase. The increase in LDL receptor in the Hver leads to an increase in uptake of LDL or HDL, with apo E, and consequently, to a greater deHvery of cholesterol to the Hver. An increase in synthesis of HMG-CoA synthase leads to an increase in synthesis of cholesterol in the Hver to meet aH ceHular needs. Thus, the presence of a CETP inhibitor in plasma wiU prevent the uptake of VLDL and/or LDL by tissues as weH as the deposition of cholesteryl esters. In general, high levels of HDL have an anti-atherogenic effect whereas high levels of LDL have an atherogenic effect. The circulation in blood of compounds, such as cholesterol, that are insoluble in water requires the formation of particles. The insoluble components, e.g., cholesteryl esters and triglycerides, are packed in the core of the particles and surrounded by polar components such as proteins, phosphoHpids and the Hke. These particles are caHed Hpoproteins, and have, thus, an outer polar sheH and a non-polar core. These associations of Hpoproteins containing CE in the core and, depending on their sizes and densities, have been named VLDL, IDL, LDL, and HDL. VLDL is the largest Hpoprotein secreted by the Hver and is converted into IDL, and then to LDL, after the triglycerides contained by the VLDL are hydrolyzed by the Hpoprotein Hpase enzyme present on the surface of the arterial walls. LDL is the major Hpoprotein that provides cholesteryl ester to extrahepatic and hepatic tissues. HDL is also secreted by the Hver and is divided in HDL2 and HDL3 on the basis of size and density. A function of HDL is to pick-up cholesterol from extrahepatic cells and deHver it to the Hver, either through VLDL and LDL or through large HDL which is enriched with apo E. These large HDL particles are caHed HDL an( i they do not sta y i tne plasma for long periods of time. They are rapidly removed by the Hver or converted back to HDL2 after donating their cholesteryl esters to VLDL and LDL. HDLi i s not present in either normal humans or non-human primates. As indicated above, HDLi appears as a distinct band in the plasma of baboons that have been fed a HCHF diet. From what is known, when cholesterol enters the blood stream it becomes associated with HDL in the form of a CE, with the help of the LCAT enzyme. In HCHF-fed baboons, this appears as an HDL -CE fraction. The CE is then transferred from HDL to VLDL and LDL to form VLDL-CE and LDL-CE with the aid of the CETP enzyme. These particles then enter the Hver ceUs through the LDL receptor (LDL-R). After being metabolized in the Hver ceUs, the VLDL-CE is returned to the plasma and thus to the periphery of the mammaHan body, where its deposition may occur leading to atherosclerosis. An inhibitor of CETP, such as the one provided by this invention, blocks the transfer of CE from HDL to VLDL and LDL. Instead, a shunt is favored that leads to the association of the CE with apo E and to the formation of HDLi-CE-apo E particles that can enter the Hver ceUs through the LDL receptor (LDL-R). The apoHpoprotein C-I of various species but not baboon, are known. The Apo C-I is a single polypeptide of molecular weight 66,000, consisting of 57 amino acids. It is a basic protein that is mainly present in VLDL and HDL, with HDL serving as a reservoir for this protein. LDL, on the other hand, contains Httle apo C-I. It has recently been shown that apo C-I displaces apo E from VLDL and affects its binding to the LDL receptor. The polypeptide inhibitor of CETP that is described herein has a common sequence with the N-termi al fragment of apo C-I. This fragment includes at least 36 amino acids as shown below. The endogenous polypeptide provided by this invention has a molecular weight of about 4,000 and becomes associated or binds to apo A- I and apo E in plasma. Its N-terminal 36 amino acids are shown below. A polypeptide having the sequence corresponding to amino acids 1 to 36 of the above sequence was synthesized by the inventors and shown to be inhibitory of CETP in vitro. The peptide has the foHowing sequence. 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ.ID.NO: 1). Fragments of that polypeptide comprising the C-terminal fragments of amino acids 28 to 36 and amino acids 16 to 36 showed limited inhibitory activity of CETP at 50 mg. However, the fragment comprising the C-terminal amino acids 28 to 36 showed at 200 mg an activity inhibitory of CETP approximately the same as that of the 36 amino acid peptide. The N-terminal fragments comprising amino acids 1 to 15, amino acids 1 to 20 and amino acids 1 to 10, as weH as the intermediate fragments comprising amino acids 15 to 30 and the Hke, corresponding to the synthesized peptide have been shown active as inhibitors of CETP. Analogues of the polypeptide of the invention and fragments thereof having inhibitory activity of CETP are also part of the invention. The analogues may have one or more substitutions in their sequences while stiH preserving their inhibitory activity. Examples of analogues suitable as inhibitors of CETP are analogues of the peptide of the invention and fragments thereof, such as those where one or more of the amino acids are substituted in accordance with the guildeHnes provided below. The substitute amino acids may be selected from the group consisting of Glu, C a -methylAsp, and β-carboxyAsp for Asp; isoVal, norVal, Leu, and C a -methylVal for Val; Gly, β -Ala, C a -methylAla, and 2-amino butyric acid for Ala; norLeu, isoLeu, and C a -methylLeu for Leu; ornithine, Arg, citrulline and C a -methylLys for Lys; Ala and 2-amino isobutyric acid for Gly; Gin, citrulline, and C a -methyl Trp for Trp; 2-amino adipic acid, Asp, and C a -methylGlu for Glu; Leu, norLeu, and C a -methylHe for He; Lys, homoArg, citrulline and C a -methylArg for Arg; Asn, citrulline, and C- a -methylGln for Gin; 2-amino-4-phenylbutyric acid, and C a -methylPhe for Phe; Ser and C a -methylThr for Thr; Thr and C a -methylSer for Ser; and 3,4-dehydroPro and C a -methylPro for Pro; and combinations thereof. However, other substituents of a nature equivalent to that of the substituted amino acid as is known in the art may also be utilized, alone or in combination with other substituents. It is therefore provided in accordance with this invention, a substantiaHy pure polypeptide having activity inhibitory of CETP. In one embodiment of the invention, the polypeptide is capable of inhibiting the binding of an about 31kD modified apo A-I polypeptide present in the plasma of high HDL baboons or a peptide of the sequence 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ.ID.NO: 1). In stiH another embodiment, the polypeptide of this invention is capable of inhibiting the binding of an about 41kD modified apo E polypeptide present in the plasma of high HDL baboons with antibody raised against the 36 amino acid N-terminal fragment of apo C-I or a peptide of the formula 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-He-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). with an antibody raised against modified apo A-I. Preferred polypeptides are the polypeptide of the sequence 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-He-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). anti-4kD peptide antibody-binding inhibitory fragments thereof; and anti-4kD peptide antibody-binding inhibitory analogues thereof having at least one substitute amino acid selected from the group consisting of Glu, C a -methylAsp, and β-carboxy Asp for Asp; isoVal, norVal, Leu, and C a -methylVal for Val; Gly, β-Ala, C a -methylAla, and 2-amino butyric acid for Ala; norLeu, isoLeu, and C a -methylLeu for Leu; ornithine, Arg, citrulline and C a -methylLys for Lys; Ala and 2-amino isobutyric acid for Gly; Gin, citrulline, and C a -methyl Trp for Trp; 2-amino adipic add, Asp, and C a -methylGlu for Glu; 2-amino adipic add, Asp, and C a -methylTrp for Trp; Leu, norLeu, and C a -methylHe for He; Lys, homoArg, dtrulline and C a -methylArg for Arg; Asn, dtrulline, and C a -methylGln for Gin; 2-amino-4-phenylbutyric add, and C a -methylPhe for Phe; Ser and C a -methylThr for Thr; Thr and c a -methylSer for Ser; 3,4-dehydroPro and C a -methylPro for Pro; and combinations thereof. In one particularly preferred embodiment, the polypeptide contains amino adds 1 through 36 of the above sequence. In stiH another particularly prefeirred embodiment, the peptide is selected from the group consisting of peptide fragments comprising amino adds 1 to 17, 1 to 20 and 1 to 25, and fragments thereof having anti-4kD peptide antibody/1-36 amino addpepti.de binding inhibitory activity. In stiH another preferred embodiment, the peptide fragments are selected from the group consisting of peptides comprising amino acids 1 to 18, and 1 to 28, and fragments thereof having anti-4kD peptide antibody/1-36 amino add peptide binding inhibitory activity. Also preferred are analogues with the Lys, Asp and Asn amino adds substituting for the Arg, Glu and Gin amino acids; the Ser, Leu, and Ala amino adds substituting for the Thr, lie and Gly amino adds; the omithine, dtrulline and aaminoadipic add amino acids substituting for the Lys and Glu amino adds. Also preferred are the foUowing analogues. Peptides comprising amino acid sequences where amide bond(s) (e.g., -C(O)-NH-) linking any pair, and up to aU pairs, of amino acids comprising amino acids 1 to 17, 1 to 20, 1 to 25, 1 to 36, and fragments thereof having anti-4kD peptide antibody/1-36 amino acid peptide binding inhibitory activity are replaced by thioether bonds (e.g., -CH2-S-), alkyl such as ethyl (-CH2-CH2-), and/or amino (e.g., -CH2-NH2) linkages. These analogues may be purchased commericaUy or prepared by methods known to those skiUed in the art as long as the antibody 1-36 amino acid peptide binding and CETP inhibitory activities of the peptides (analogues) are retained to some degree. However, other analogues are also part of this invention as long as they preserve the inhibitory activity of the antibody/ 1-36 amino acid peptide binding. The CETP inhibitory polypeptide of the invention may be provided as a powder, preferably in freeze-dried form, as a solution, preferably frozen at below -20 °C, and the Hke, to prevent proteolysis. This invention also provides an anti-atherosclerosis composition, comprising an anti-atherosderosis effective amount of the polypeptide of the invention; and a pharmaceuticaHy-acceptable carrier. When the composition is used as a preventative tool, it may contain 10 to 200 mg and more preferably 20 to 100 mg of the polypeptide. However, other amounts are also suitable. When the composition is intended for therapeutic use, the amount of polypeptide present is preferably about 10 to 400 mg, and more preferably about 20 to 300 mg. However, other amounts may also be utilized. Any and all pharmaceuticaHy-acceptable carriers known in the art for administration of peptides to mammals, and preferably to humans, are suitable for use herein. These are known in the art and need not be further described herein. Examples, however, are saline, human serum albumin and starch. However, others may also be utilized. The composition of this invention may be provided in unit form, preferably in a sterile, dosed container, and more preferably in a sealed container. A kit, comprising in separate containers as least one unit of the anti-atherosderosis composition of the invention; at least one syringe; and at least one needle. TypicaHy, a kit may contain from about 1 to 20 units of the composition of the invention, but could contain 50 units or more. In addition, the Ht may contain 1 to 20, but and sometimes 50 or more syringes if they are disposable, and 1 to 20, but sometimes up to 50 or more needles if they are disposable. The components of the kit are provided in a sterile form, be it wrapped in a sealed, sterilized wrapping, or in some other way. If not disposable, the syringe and needle may be autodaved between uses. The composition of the invention is preferably administered intravenously, although it may also be administered intraperitoneaUy, subcutaneously or intramuscularly. The oral route is not permissible since the polypeptide would be degraded in the addic pH of the stomach. The composition may preferably have a pH of about 7 to 9, and more preferably about 8 to 9, which may be adjusted with the addition of a base, add or buffer as is known in the art. This invention also provides an antibody having specificity for a polypeptide selected from the group consisting of the polypeptide of the invention; the baboon CETP polypeptide inhibitor, fragments thereof and analogues thereof; the 1-36 amino acid N-terminal fragment of apo C-I; modified apo A-I (MW:31kD); and modified apo E (MW:41kD). The antibodies of the invention may be raised in mammals as is known in the art. (Albers, J.J., Hazzard, W.R., "Immunochemical Quantification of the Human Lp(a) Lipoprotein", Lipids 9:15-26(1974)). TypicaUy, the antibodies may be raised in rabbit, goat, sheep, pig and chicken. However, other mammals may also be utilized. Preferred are rabbit antibodies. Also preferred are polyclonal antibodies. However, monoclonal antibodies may also be prepared by methods known in the art (Kohler, G., and MHstein, D., "Continuous Cultures of Fused CeUs Secreting Antibody of Predefined Specificity", Nature (London) 256:495-497(1975)). In one preferred embodiment, the antibody of the invention is capable of specificaUy binding to modified apo A-I. In another preferred embodiment, the antibody is capable of specificaUy binding to the baboon CETP inhibitor polypeptide of this invention. In stiH another embodiment, the antibody of the invention is capable of specificaUy binding to the polypeptide of the sequence 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-ne- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1), fragments thereof and analogues thereof as described above. In stiH another preferred embodiment of the invention, the antibody is capable of specificaUy binding to modified apo E. The antibody of the invention, in another preferred embodiment is also capable of specificaUy binding to apo C-I, and more preferably to the 1-36 N-terminal fragment thereof. In another aspect of the invention, a method is provided for preventing atherosderosis in a mammal being predisposed to that condition. The method comprises administering to the mammal a prophylacticaHy effective amount of the polypeptide of the invention, or of the anti-atherosderosis composition described above. In a preferred embodiment, the polypeptide is administered in an amount of about 10 to 200 mg, and preferably about 2 to 100 mg for preventative appHcations. However, other amounts may also be administered. The polypeptide or composition thereof may be aό ninistered in a smaU volume of carrier, e.g., 0.2 to 1.5 ml of saline or other carriers, as is known in the art. The polypeptide of the invention may be administered intravenously, to a fragment of the population, particularly the human population, that is not afflicted by high blood cholesterol and hyperbetaHpoproteinemia, but, as determined by other means, may be at risk of being afflicted by atherosclerosis. One such example may be a familial trait having been determined. The polypeptide of this invention may be administered on a daUy basis, or at longer intervals if provided as a slow release composition as is known in the art, such as depoestradiol provided by the UpJohn Co. (Up ohn Co., Kalamazoo, MI). In another aspect, the present invention provides a method of treating a mammal afflicted with atherosclerosis. The method comprises administering to the mammal a therapeuticaUy affective amount of the polypeptide of the invention. When administered for therapeutic purposes, the polypeptide may be injected in an amount of about 10 to 400 mg, and more preferably 20 to 300 mg. However, other amounts as assessed by a practitioner in specific cases, may also be administered. In this case, as in the case of the prophylactic administration, the polypeptide may be administered intravenously, among other routes. Having now generaUy described this invention, the same wiU be better understood by reference to certain specific examples, which are included herein for purposes of Ulustration only and are not intended to limiting of the invention or any embodiment thereof, unless so specified. EXAMPLES Example 1 : Animals and Diet Adult male and female baboons (papio sp.) held in a baboon colony at the Southwest Foundation for Biomedical Research in San Antonio, Texas, were used as blood donors for these studies. Among these, 24 baboons had a high HDLi phenotype and 32 had a low HDLi phenotype (WilHams, M.C., et al, "Detection of abnormal Hpoprotein in a large colony of pedigreed baboons using high-performance gel exclusion chromatography", J. Chromat. 308:101-109(1984)). Half of the high HDL baboons (n=16) were maintained on a HCHF diet, the composition of which has been previously described (Kushwaha, R. S., et al, "MetaboHsm of apoHpoprotein B. in baboons with low and high levels of low density Hpoproteins", J. Iipid Res. 27:497-507(1986)). Most of the baboon donors of the low HDL phenotype were maintained on a chow diet (Purina Monkey Chow, manufactured by Ralston Purina Co., St. Louis, Missouri). The monkey chow is low in fat (10% of total calories) and high in carbohydrate (62% of total calories). In addition, the chow has a very low cholesterol content (0.03 mg/Kcal). The high HDLi baboons were progeny of two sires (X1672 and X102) who had a high HDLi phenotype. The low HDL baboons were progeny of a number of sires who did not have a high HDLi phenotype. The presence of HDL was detected by high performance Hquid chromatography (HPLC) as described previously (Williams et al "Detection of abnormal Hpoprotein in a large colony of pedigreed baboons using high-performance gel exdusion chromatography", J. Chromatography 308:101-109(1984)). Example 2; Preparation of *--H Cholesteryl Ester HDL High and low HDLi baboons were immobilized with 10 mg/kg of ketamine HC1 and bled. The blood was coUected in tubes containing 1 mg/ml EDTA, and plasma separated by low speed centrifugation at 6 °C. The plasma was treated with sodium azide, chloramphenicol, gentamydn sulfate, phenylmethyl-sulfonyl fluoride and DTNB as described previously (Kushwaha, R. S., et al, "Impaired plasma CE transfer with accumulation of larger high density Hpoproteins in some families of baboons (Papio sp.)," J. Lipid Res. 31:965-973(1990)). 20 to 60 mCi of tritiated cholesteryl linoleate were dissolved in ethanol and then added to the plasma. The plasma was flushed with nitrogen and incubated for 20 hrs at 4 °C. After incubation, HDL3 was isolated by density gradient ultracentrifugation (McGill, H. C, et al, "Dietary effects on serum Hpoproteins of dysHpopro-einemic baboons with high HDL ", Arteriosclerosis 6:651-663(1986); Redgrave, T. G., et al, "Separation of plasma Hpoproteins by density-gradient ultracentri-fugation", AnaLBiochem. 65:42-49(1975)), dialyzed against saline/EDTA, and used as a substrate for the CE transfer reaction. The total and free cholesterol contents of HDL3 were measured prior to use in the assay. Example 3: Preparation of Acceptor Lipoproteins and CETP Source VLDL+LDL from low HDLi baboons was used as the acceptor of CE from HDL3. A VLDL+LDL fraction of d<1.040 g/ml was isolated from 100-200 ml of blood by sequential ultracentrifugation as described previously (Kushwaha et al (1986), supra). Total and free cholesterol contents in acceptor Hpoproteins were measured by enzymatic methods (Wako Pure Chemical Co.) (AHain, CC, et al., "Enzymatic Determination of Total Serum Cholesterol", Clin. Chem. 20:470-475(1974). After separation of VLDL+LDL, the bottom fraction was adjusted to d=1.21 g/ml by adding soHd KBr, and total Hpoproteins were isolated by ultracentrifugation (Kushwaha et al (1986), supra). The bottom fraction of d 1 1.21 g/ml was also coUected. AU Hpoprotein fractions and the Hpoprotein-defident fraction of d 1 1.21 g/ml (LPDS) were dialyzed against saline/EDTA. The LPDS was used as the source of CETP. Example 4: Cholesteryl Ester Transfer Assay The CE transfer activity of a sample was assayed by a modification of a procedure described previously (Kushwaha et al, (1986), supra). Briefly, _ 3 H CE-labeled HDL3 containing50-100 mg of CE from low HDLi baboons was incubated with VLDL+LDL containing 100-300 mg CE in the presence of LPDS. The acceptor Hpoproteins and the LPDS were obtained from low HDLi baboon plasma (chow diet). In some cases, the HDL3 was obtained from high HDLi baboons maintained on the HCHF diet. The incubations were carried out at 4 °C (control) and 37 °C for 4-6 hrs, and terminated by placing the samples on ice. The assay mixture was then ultracentrifuged to separate VLDL+LDL having a ά| 1.040 g/ml, and the radio-activity in the Hpoproteins was counted as described previously (Kushwaha et al, (1986), supra). Any difference observed in the radioactivity transferred from HDL to VLDL+LDL at 4 °C and 37 °C was attributed to CETP activity in the LPDS. Time course experiments gave a linear response up to 7 hrs. Similarly, the CETP activity was linear with increasing LPDS, up to 140 ml LPDS, which was derived from an equivalent volume of plasma. The polypeptide of the invention, and synthetic fragments thereof were added to the reaction mixture to determine their CETP inhibitory activities, along with an appropriate synthetic control peptide. The percent difference between the control experiment and the assay with inhibitor peptide was expressed as the inhibitor activity. Example 5: Identification of Inhibitor Polypeptide The bottom non-Hpoprotein fraction obtained by ultracentrifugation for 72 hrs was analyzed for protein content by 10 % SDS-polyacrylamide gel electrophoresis (LaemmH, U. K., "Cleavage of structural proteins during the assembly of the head of bacteriophage T4", Nature 227:680-685(1970)), without β-mercaptoethanol pretreatment. To determine differences in smaH molecular weight apoHpoproteins, deHpidated Hpoproteins of d<1.21 g/ml from high and low HDL baboons were separated by 15-19% SDS-polyacrylamide gel electrophoresis with β-mercapto-ethanol pretreatment prior to loading the samples onto the gels. Example 6: Electroelution of Inhibitor Polypeptide The Hpoproteins fraction of d<1.21 g/ml from high HDLi baboon plasma was deHpidated with ether:ethanol (Floren, C.H., et al, "Estrogen-induced increase in uptake of cholesterol-rich very low density Hpoproteins in perfused rabbit Hver", MetaboHsm 30:367-374(1981)), and separated by 15% SDS gel electrophoresis (LaemmH, (1970), supra) after addition of β-mercaptoethanol. A smaH molecular weight protein band was cut and transferred onto a tube gel (15%). A dialysis tube of molecular weight cut-off point 1000 was attached to the bottom of the gel tube to receive the electroeluted peptide. The thus electroeluted peptide was dialyzed and quantitated by comparing its absorbance at 660 nm with a known amount of stained albumin electroeluted at the same time. Example 7: Antibody Preparation The apoHpoproteins were separated by 15% SDS-gel electrophoresis and stained (LaemmH, (1970), supra). The stained bands were transferred onto a nitroceUulose membrane. The bands corresponding to the inhibitor polypeptide were cut out (0.05 mg) and dissolved in 0.5 ml of filtered DMSO. 0.5 ml Freund's adjuvant were then added and thoroughly mixed and the mixture was injected intradermaUy into two rabbits. After 30 days, the rabbits were boosted with a similar amount of electroeluted protein band. Antibody titer was measured by Western blotting. The rabbits were boosted again 3 times. For the preparation of antibody against the synthetic polypeptide, 500 mg of polypeptide were dissolved in 400 ml of titer max (CytRx Corporation, Atlanta, GA), and injected into rabbits intradermaUy. The rabbits were boosted with 500 mg of the synthetic peptide in 200 ml of Titer Max on the 28th day. The serum was tested on the 42nd day. The rabbits were bled and anti-serum was obtained as needed. Example 8: Immunoaffinity Chromatography An immunoaffinity column was prepared using CnBr-activated Sepharose beads (Pharmada Co.). The bound Hgand was IgG precipitated from the serum of rabbits having antibodies. The method used to predpitate IgG was similar to that described by McKinney and Parkinson (McKinney, M. M. and Parkinson, A., "A simple, non-chromatographic procedure to purify i m unoglobins from serum and asdtes fluid", J. Tmmunological Methods, 96:271-278(1987)). Briefly, 5 ml of rabbit serum were dUuted 4-fold with acetate buffer, pH 4.0. 625 ml of capryHc add were added dropwise to precipitate albiimin and non-IgG proteins. The insoluble materials were removed by centrifugation at 10,000xg for 30 min. The supernatant was mixed with phosphate buffered saline, and the pH adjusted to 7.4 with IN sodium hydroxide. The solution was cooled to 4 °C and ammonium sulfate was added to give a final concentration of 45% to predpitate the IgG. The precipitate was recovered as a peUet after centrifugation, and resuspended in phosphate buffered saline. The IgG was dialyzed overnight in 100 volumes of phosphate buffered saline, and then dissolved in sodium acetate buffer, pH 8.3, coupled to 3 g of CnBr-activated Sepharose beads, and maintained in Tris-saHne, pH 7.4, until ready to use. 8 ml of plasma were incubated overnight with IgG coupled beads in Tris-saHne buffer with gentle rotation. The column was then washed with Tris-saHne coupHng buffer and sodium acetate buffer as described by Cheung and Albers (Cheung, M.C., and Albers, "Distribution of high density Hpoprotein particles with different apoHpoprotein composition: particles with A-I and A-II and particles with A-I but no A-II", J. Lipid Res. 23:747-753(1982)). The bound proteins were eluted with 0.1 M acetic acid, pH 3.0, and 1 ml aHquots were coUected and read at 280 nm to visualize the peak. The protein fraction was dialyzed immediately against phosphate buffered saline and separated by electrophoresis in 15% SDS-polyacrylamide gels. Example 9: Immunoblotting The proteins separated with SDS-polyacrylamide gels were transferred onto ImmobUon-P sheets (MilHpore, Bedford, MA). The sheets were incubated with antibody against inhibitor peptides after blocking of nonspecific sites. The sheets were washed and incubated again with a secondary antibody containing horseradish peroxidase. The addition of boric acid buffer containing 3-amino-9-ethylcarbazole, methanol and hydrogen peroxide produced a coloration. Example 10: Amino Add Analysis and Sequendng Stained bands of proteins were transferred onto TmmobUon-P sheets. Selected bands were cut out and hydrolyzed with 50% propionic add, 50% 12 N HC1 for 2 hrs at 135 °C. Amino acid analysis of each sample was performed using a model 6300 amino add analyzer (Beckman Co, Palo Alto, CA) provided with System Gold software. The same bands were sequenced using a model 477A protein sequencer (AppHed Biosystems, Foster City, CA). Example 11: Preparation of Synthetic Peptides The peptides were synthesized by soHd-phase peptide synthesis as described by Barany and Merrifield (Barany, G., and Merrifield, R. B., "The Peptides", Analysis, Synthesis, Biology, Gross, E. and Meienhofer, J., eds., Vol 2, Academic Press, New York, pp. 1-284(1980)). The 1-36 amino add synthetic peptide was assembled from the C-terminus towards the N-terminus, with the a-carboxyl group of the amino add attached to a soHd support and was then characterized by HPLC. Example 12: Data Analysis The values provided in the foUowing examples are averages and are provided as mean + standard error. These values were compared using variance analysis and, if significant differences were detected, the values were compared using Duncan's Multiple Range Test (Duncan, D.B., "Multiple Range and Multiple F Tests", Biometrics 11: 1-12 (1955) ) . Example 13: Characterization of Proteins from Infranatant Fraction The CETP inhibitory activity was lost from the HDL when the Hpoproteins were extensively ultracentrifuged, e.g., for 72 hrs, or by repeated ultracentrifugation. After ultracentrifugation, the inhibitory activity was found in the infranatant fraction. To characterize the proteins, the infranatant fraction (d<1.21 g/ml) was separated by 10% SDS polyacrylamide gel electrophoresis in the absence of β-mercaptoethanol. The infranatant fraction from high HDL baboons contained albun in, a protein sHghtly larger than apo A-I and another protein larger than apo E. Lanes A and B show nonHpoprotein fractions from high and low HDLi baboons, respectively. The protein bands 1, 3, and 5 correspond to albumin, apo E and apo A-I, respectively. The protein bands 2 and 4 of molecular weights between 1 and 3, and 3 and 5, respectively, correspond to proteins with molecular weights of 41,000 and 31,000, respectively. Protein samples from high HDLi baboons show only bands corresponding to albumin, a protein of molecular weight 41,000, and a protein of molecular weight 31,000. The molecular weights were determined with standard proteins separated on si Uar gels (gel picture not shown). The infranatant fraction from low HDLi baboons contains these proteins as weH, but in addition, it also contains apo A-I and apo E. Both proteins in the apo A-I region were identified by immunoblotting with antibody to apo A-I. Similarly, both proteins in the apo E region were identified by immunoblotting with antibody to apo E. The molecular weights of the proteins detected by immuno-blotting with apo A-I and apo E show a difference of about 4 kD (picture not shown). The estimated molecular weight of apo A-I is about 27,500 and that of the modified apo A-I is about 31,000. Similarly, the estimated molecular weight of apo E is about 37,000 and that of modified apo E is about 41,000. Both apo A-I and apo E are modified by a protein of about 4,000 molecular weight. Example 14: Detection of CETP Inhibitor Peptide in Plasma of High HDLi Baboons To determine if a common polypeptide of molecular weight 4,000 was modifying both apo A-I and apo E, plasma Hpoproteins of d<1.21 g/ml were separated from high and low HDLi baboons by 18% SDS-polyacrylamide gel electrophoresis with β-mercaptoethanol. A higher amount of a 4kD protein was detected in Hpoprotein from high HDLi baboons as compared to Hpoprotein from low HDL baboons (gel picture not shown). To determine if the 4kD polypeptide inhibits CE transfer, the polypeptide and albumin were electroluted from the gels, and used in increasing concentrations in a CE transfer assay mixture with Hpoproteins from low HDL baboons as described in Example 4 above. Albumin had no effect on the transfer of CE from HDL to VLDL+LDL. The 4kD polypeptide, on the other hand, significantly inhibited CETP activity. (Results are not shown). Example 15: Characterization of Polypeptide by Affinity Chromatography Rabbit antibody was prepared against the 4kD polypeptide isolated from Hpoproteins obtained from high HDLi baboons. This antibody was used to prepare an immunoaffinity column. The Hpoproteins of d<1.21 g/ml were passed over the immunoaffinity column. The bound Hpoproteins eluted with OJ M acetic add and separated in 15% SDS-polyacrylamide reducing gels. 4kD, and 31kD poly-peptides, and a minor band corresponding to a 41 kD polypeptide were detected. The column bound peptides (100 mg) inhibited by 31.3 + 1.4% the transfer of CE (mean ± SE, n=3) in the CETP assay from low HDLi baboons. On the other hand, the addition of IgG to the assay from high HDLi baboons increased CE transfer by 44.3 + 1.5 % (n=3), but had no effect on CE transfer from low HDLi baboons. Example 16: Comparison of 4kD Inhibitor Polypeptide with Sequence Data Bank The sequence of this polypeptide was compared to sequences of known proteins using Sequence Data Bank (Reardon, W. R. and Lipman, D. J., PNAS (USA) 85:2444-2448(1988)), and was found to have 100% homology with human and crab-eating macaque apo C-I. The sequence was then compared with apo C-I from baboons and found to be 100% homologous (private communication from Dr. Hixson of the Southwest Foundation for Biomedical Research). Example 17: Characterization of Inhibitor Polypeptide with Synthetic Peptides Based on its molecular weight, it was determined that the 4kD inhibitor polypeptide contained approximately 36 amino acids. Three peptides were synthesized beginning from the C-terminal end of the apo C-I sequence. The first peptide contained 9 amino acids, the second peptide contained 21 amino acids and the third peptide contained 36 amino acids. The 36 amino acid peptide had an amino acid sequence similar to the 4,000 MW polypeptide , the other two were fragments of the synthetic peptide starting from its C-terminus. 50 mg of these peptides were used in a CETP assay with Hpoproteins of low HDLi baboons as described in Example 4 above. A soluble heHcal peptide with a 1,900 molecular weight utilized as control. The 36 amino acid polypeptide significantly (p<0.01) inhibited CE transfer from HDL to VLDL and LDL, whUe the others, induding the control peptide, did not. Example 18: Antibody Against 36 Amino Add Inhibitor Peptide Antibody against the 36 amino add inhibitor peptide was prepared in rabbits as described in Example 7, and used for immunoblotting. The thus prepared antibody recognized the 4kD peptide as weH as a 31kD polypeptide from Hpoproteins of high HDLi baboons. To determine if both the apo C-I and the 4kD polypeptide were present in the plasma of high and low HDLi baboons, Hpoproteins from both phenotypes were separated by 10% SDS gel electrophoresis and immuno-blotted. Two protein bands were detected with the antibody by imm un oblotting of samples from high HDL baboons. Only a single band was detected in samples from low HDL baboons. In addition, isoelectric focusing patterns of the synthetic peptide suggest that the peptide is a sHghtly basic protein. Example 19: CETP Inhibition by Various Peptide Fragments A CE transfer assay as described in Example 4 above was conducted using HDLi baboon plasma, ^H HDL, and the VLDL+LDL carriers, in the presence of CETP enzyme to mediate the exchange. The reactions were conducted at 37 °C and at 4 °C (control) in dupHcate. The results are shown in Table 3 below. Table 3: CETP Inhibition by Various Peptide Fragments CETP [H3] HDL Synthetic VLDL+LDL Temp. Inhibition Peptide 125 100 300

Claims: CLAIMS 1. A substantiaUy pure polypeptide having activity inhibitory of CE transfer protein (CETP). 2. The CETP inhibitory polypeptide of daim 1, that is capable of inhibiting the binding of an about 31kD modified apo A-I polypeptide present in the plasma of high HDLi, baboons or a peptide of the sequence 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-ne-Lys-Ghi-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ..ID.NO: 1), to an antibody raised against the above peptide. 3. The CETP inhibitory polypeptide of daim 1, that is capable of inhibiting the binding of an about 4 kD CETP inhibitor polypeptide present in the plasma of high HDL baboons to an antibody raised against a peptide of the formula 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ue- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-ϋe-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). 4. The CETP inhibitory polypeptide of claim 1, that is capable of inhibiting the binding of an about 41kD modified apo E polypeptide present in the plasma of high HDLi baboons with an antibody raised against the N-terminal fragment of apo C-I or a peptide of the formula 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). 5. The CETP inhibitor polypeptide of claim 1, that is capable of inhibiting the binding of the N-terminal fragment of apo C-I or a peptide of the formula 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1), with an antibody raised against apo A-I. 6. The CETP inhibitory polypeptide of daim 1, comprising a peptide selected from the group consisting of the amino acid sequence 1 2 3 4 5 6 7 8 9 10 11 12 Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- 13 14 15 16 17 18 19 20 21 22 23 24 Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-ne- 25 26 27 28 29 30 31 32 33 34 35 36 Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ.ID. NO: 1), anti-4kD peptide antibody-binding inhibitory fragments thereof; and anti-4kD peptide antibody-binding inhibitory analogues thereof having at least one substitute amino add selected from the group consisting of Glu, C a -methylAsp, and β-carboxy Asp for Asp; iso Val, norVal, Leu, and C a -methylVal for Val; Gly, β-Ala, C a -methylAla, and 2-amino butyric add for Ala; norLeu, isoLeu, and C a -methylLeu for Leu; omithine, Arg, dtrulline and C a -methylLys for Lys; Ala and 2-amino isobutyric add for Gly; Gin, citiruUine, and C a -methyl Asn for Asn; p-benzoyl Phe and C a -methyl Trp for Trp; 2-amino adipic acid, Asp, and C a -methylGlu for Glu; Leu, norLeu, and C a -methyllle for lie; Lys, homoArg, dtrulline and C a -methylArg for Arg; Asn, dtrulline, and C- a -methylGln for Gin; 2-amino-4-phenylbutyric add, and C a -methylPhe for Phe; Ser and C a -methylThr for Thr; Thr and C a -methylSer for Ser; and 3,4-dehydroPro and C a -methylPro for Pro; sequences where at least one amino bond is replaced by thioether, alkyl or alkyl amino; and combinations thereof. 7. The CETP inhibitory polypeptide of claim 6, wherein the peptide comprises amino acids 1 to 36 of the amino acid sequence. 8. The CETP inhibitory polypeptide of daim 6, wherein the peptide is selected from the group consisting of peptide fragments comprising amino acids 1 to 17, 1 to 20, 1 to 25, and fragments thereof having anti-4kD peptide antibody/1-36 amino acid peptide binding inhibitory activity. 9. The CETP inhibitory polypeptide of claim 7, wherein the peptide fragments are selected from the group consisting of peptides comprising amino acids 1 to 18, 1 to 28, and fragments thereof having anti-4kD peptide antibody/1-36 amino acid peptide binding inhibitory activity. 10. The CETP inhibitory polypeptide of claim 1, in freeze dried form. 11. An anti-atherosclerosis composition, comprising an anti- atherosclerosis effective amount of the polypeptide of claim 1; and a pharmaceuticaUy-acceptable carrier. 12. The anti-atherosderosis composition of claim 1, comprising about 10 to 400 mg of the polypeptide. 13. The anti-atherosclerosis composition of claim 1, in unit form. 14. An anti-atherosclerosis kit, comprising in separate sterile containers at least one unit of the composition of claim 11; at least one s;j * ringe; and at least one needle. 15. An antibody having specifidty for a polypeptide selected form the group consisting of the polypeptide of daim 1; baboon CETP polypeptide inhibitor; 1-36 amino add N-terminal fragment of apo C-I; modified apo A-I (MW:31kD) ; and modified apo E (MW:41kD). 16. The antibody of daim 15, being a polydonal antibody. 17. The antibody of daim 15, being capable of specificaUy binding to modified apo A-I. 18. The antibody of daim 15, being capable of specificaUy binding to the baboon CETP inhibitor polypeptide. 19. The antibody of daim 15, being capable of specificaUy binding to modified apo E. 20. The antibody of daim 15, being capable of specificaUy binding to apo C-I. 21. The antibody of daim 15, being capable of specificaUy binding to the polypeptide. 22. A method of preventing atherosderosis in a mammal being predisposed to that condition, comprising administering to the mammal a prophylacticaHy effective amount of the polypeptide of daim 1. 23. The method of daim 22, wherein the polypeptide is administered in an amount of about 10 to 200 mg. 24. The method of claim 22, wherein the polypeptide is administered intravenously. 25. The method of claim 22, wherein the mammal is a human. 26. A method of treating a mammal afflicted with atherosclerosis comprising administering to the mammal a therapeuticaUy effective amount of the polypeptide of claim 1. 27. The method of claim 26, wherein the polypeptide is administered in an amount of about 10 to 400 mg. 28. The method of claim 26, wherein the polypeptide is administered intravenously. 29. The method of daim 26, wherein the mammal is a human. AMENDED CLAIMS [received by the International Bureau on 4 May 1993 (04.05.93); original claims 2-5,7,9,12,13 and 21 deleted; original claims 1,6,8,10,11,14,15,22 and 26 amended; remaining claims unchanged (6 pages)] 1. A substantiaUy pure polypeptide having activity inhibitory of cholesteryl ester transfer protein comprising at least a part of sequence Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- Asn-Arg-Ile-Lys-GHi-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO:l). 6. The cholesteryl ester transfer protein inhibitory polypeptide of daim 1, having at least one substitute amino add or amino bond; wherein the substitute amino add for Asp is selected from the group consisting of Glu, C a -methylAsp, and β-carboxy Asp; the substitute amino add for Val is selected from the group consisting of isoVal, norVal, Leu, and C a -methylVal; the substitute amino add for Ala is selected from the group consisting of Gly, β-Ala, C a -methylAla, and 2-amino butyric acid; the substitute amino add for Leu is selected from the group consisting of norLeu, isoLeu, and C a -methylLeu; the substitute amino add for Lys is selected from the group consisting of omithine, Arg, dtrulline and C a -methylLys; the substitute amino add for Gly is selected from the group consisting of Ala and 2-amino isobutyric add; the substitute amino add for Asn is selected from the group consisting of Gin, dtrulline, and C a -methyl Asn; the substitute amino add for Trp is selected from the group consisting of p-benzoyl Phe and C a -methyl Trp; the substitute amino acid for Glu is selected from the group consisting of 2-amino adipic acid, Asp, and C a -methylGlu; the substitute amino acid for lie is selected from the group consisting of Leu, norLeu, and C a -methylfle; the substitute amino add for Arg is selected from the group consisting of Lys, homoArg, citrulline and C a -methylArg; the substitute amino add for Gin is selected from the group consisting of Asn, citrulline, and C- a -methylGln; the substitute amino acid for Phe is selected from the group consisting of 2-an ino-4-phenylbutyric add and C a -methylPhe; the substitute amino acid for Thr is selected from the group consisting of Ser and C a -methylThr; the substitute amino add for Ser is selected from the group consisting of Thr and C a -methylSer; the substitute amino add for Pro is selected from the group consisting of 3,4-dehydroPro and C a -methylPro; and the substitute amino acid bond is selected from the group consisting of thioether, alkyl alkyl amino, and combinations of thioether, alkyl and alkyl amino bonds. 8. The cholesteryl ester transfer protein inhibitory polypeptide of daim 1, wherein the part of the peptide sequence is selected from the group consisting of amino adds 1 to 17, amino adds 1 to 20 and amino adds 1 to 25. 10. The cholesteryl ester transfer protein inhibitory polypeptide of daim 1, in freeze dried form. 11. An anti-atherosderosis composition, comprising an anti-atherosderosis effective amount of at least a part of a polypeptide of of the amino add sequence Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ.ID. NO: 1); and a pharmaceuticaHy-acceptable carrier. 14. An anti-atherosderosis kit, comprising in separate sterile containers at least 10 to 400 mg of the composition of daim 11; at least one syringe; and at least one needle. 15. An antibody having spedfidty for a polypeptide comprising at least a part of a peptide of sequence Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Gal-Ile- Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). 16. The antibody of claim 15, being a polyclonal antibody. 17. The antibody of daim 15, being capable of specificaUy binding to modified apo A-I. 18. The antibody of daim 15, being capable of specificaUy binding to the baboon CETP inhibitor polypeptide. 19. The antibody of daim 15, being capable of specificaUy binding to modified apo E. 20. The antibody of claim 15, being capable of specificaUy binding to apo C-I. 22. A method of preventing atherosderosis in a mammal being predisposed to that condition, comprising administering to the mammal a prophylacticaHy effective amount of a polypeptide comprising at least a part of a peptide of sequence Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-lle- Asn-Arg-Ile-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). 23. The method of daim 22 , wherein the polypeptide is administered in an amount of about 10 to 200 mg. 24. The method of daim 22, wherein the polypeptide is administered intravenously. 25. The method of daim 22, wherein the mammal is a human. 26. A method of treating a mammal afflicted with atherosderosis comprising administering to the mammal a therapeuticaUy effective amount of a polypeptide comprising at least a part of sequence Asp-Val-Ser-Ser-Ala-Leu-Asp-Lys-Leu-Lys-Glu-Phe- Gly-Asn-Thr-Leu-Glu-Asp-Lys-Ala-Trp-Glu-Val-Ile- Asn-Arg-ne-Lys-Gln-Ser-Glu-Phe-Pro-Ala-Lys-Thr (SEQ. ID. NO: 1). 27. The method of daim 26, wherein the polypeptide is administered in an amount of about 10 to 400 mg. 28. The method of claim 26, wherein the polypeptide is ao -oinistered intravenously. 29. The method of daim 26, wherein the mammal is a human.

Inventors: BORN KATHLEEN, DUNHAM RAYMOND G, KANDA PATRICK, KUSHWAHA RAMPRATAP, MCGILL HENRY C JR, BORN, KATHLEEN, DUNHAM, RAYMOND, G., KANDA, PATRICK, KUSHWAHA, RAMPRATAP, MCGILL, HENRY, C., JR.
Applicants: SOUTHWEST FOUND BIOMED RES, SOUTHWEST FOUNDATION FOR BIOMEDICAL RESEARCH
Application year: 1992
Main classification: A61K37
IPC codes: A61K38, C07K14, C07K16