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`TIPS — November 1987 [Vol. 8]
`
`Advisory Editorial Board
`W. N. Aldridge
`Carshalton, UK
`E, I. Ari'e'ns
`Nijmegen, The Netherlands
`H. R. Besch Ir
`lndiar1apolis,lN, USA
`A. M. Breckenridge
`Liverpool, UK
`'1”. F. Burks
`Tucson, AZ, USA
`I. I. Burns
`New York, NY, USA
`A, W. Cuthbert
`Cambridge, UK
`C. T. Dollery
`London, UK
`I, R. Fozard
`syrasbourg, France
`T, Godfraind
`Lguvain, Belgium
`1". Greengard
`New York, NY, LISA
`15, Habermann
`Giessen, PRC
`]_ W. Hadden
`Tampa, FL, USA
`3, I-lolmstedt
`Stockholm, Sweden
`M, C. Horning
`Houston, TX, USA
`E. Hosoya
`‘I‘okyo./HIM"
`I, Hughes
`Cambridge, UK
`G, L. Plaa
`Montreal, Canada
`A, Pletscher
`Basle, Switzerland
`M, J. Rand
`Parkville, Vic, Australia
`M, M. Reideriberg
`Ngw York, NY, USA
`G, A. Robison
`Houston, TX, USA
`F. Sjoqvist
`Huddinge, Sweden
`1:, J, E. Stefano
`guenas Aires, Argentina
`U, Schwabe
`Heidelberg, FRO
`p, Taylor
`L, /glla, CA, USA
`H, Timmerman
`Amgterdam, 'I'lie Netherlands
`N, Weiner
`pgnver, CO, USA
`'1‘, C. Westfall
`5;, Louis, MO, USA
`C, Zbinden
`Zurich, Switzerland
`
`CONTENTS
`
`New insights into the non-nicotine regulation of adrenal medullary
`function, Philip D. Marley
`
`Drug Regulatory News
`
`Current Awareness
`
`MK-801, NMDA receptors and ischaemia-induced neurodegeneration,
`I. A. Kemp, A. C. Foster, R. Gill and G. N. Woodruff
`Remembering the gut—brain connection, Carlos A. Netto and
`Ivan Izquierdo
`
`This and that: on books and belle donne, antimony and anti-caffeine, B. Max
`
`Letters
`
`D1 and D2 dopamine agonist synergism and the nucleus accumbens,
`David M. Iackson, Kaba Dreher and Svante B. Ross
`Pharmacology and statistics: the Bloody Oblivious tester, A. R. Unwin
`Ian Kitchen replies
`
`Comment
`Alternative molecular interpretations of binding Curves:
`compelling competition,
`Antonio DeBlasi and Harvey I. Motulsky
`
`Viewpoint
`Agonists, partial agonists, antagonists, inverse agonists and
`agonist/antagonists?, Terry Kenakin
`
`TiPS Reviews
`
`‘
`
`Muscarinic cholinergic receptor structure: molecular biological support
`for subtypes, Anthony R. Kerlavage, Claire M. Fraser and
`I. Craig Venter
`5-I-ITIA receptor—related anxiolytics, Iiirg Traber and Thomas Glaser
`Histamine and its lymphocyte-selective derivatives as immune
`modulators, Kenneth L. Melmon and Manzoor M. Khan
`Synthesis, SARs and therapeutic potential of HMG-CoA reductase
`inhibitors, Ta-Iyh Lee
`HUMANS V. ANIMALS: Effects of 01- and [3-adrenergic agonists,
`phosphodiesterase inhibitors and adenosine on isolated human heart
`muscle preparations,
`Wilhelm Schmitz, Hasso Scholz and Erland Erdmann
`
`Editor A. C. Abbott
`Journal secretary Carol Smith
`production Sue Perkins
`Elsevier Publications Cambridge
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`Book Reviews
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`Pharmacology, by H. P. Rang and M. M. Dale
`Steroids in the nuclear zone, edited by C. R. Clark
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`Diary
`
`Index to Advertisers
`
`IX
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`Mylan Exhibit 1013, Page 2
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`442 Synthesis, SARs and therapeutic potential of HMG- CoA reductase inhibitors Ta-Jyh Lee Elevated plasma levels of low-density lipoprotein cholesterol is a major risk factor for the development of coronary heart disease, the leading cause of death and disability in Western countries. Because most cholesterol in the body is synthesized de novo, the control of endogenous cholesterogenesis would be an attractive and potentially effective means of lowering plasma cholesterol levels. This approach has been validated by the recent discoveries of two novel fungal metabolites, mevastatin and Iovastatin. These compounds are potent inhibitors of HMG-CoA reductase, which regulates the rate-limiting step in the biosynthetic pathway of cholesterol. Ta-Jyh Lee discusses the rationale for the design and synthesis of several potent inhibitors related to mevastatin and Iovastatin, and the therapeutic potential of HMG-CoA reductase inhibitors. TIPS - November 1987 [Vol. 8] practical realization of this goal has been advanced greatly by the recent discoveries of several novel fungal metabolites, which are potent inhibitors of HMG-CoA reductase. Inhibitory activities of mevastatin and lovastatin Mevastatin (Fig. 2,1a, formerly known as ML-236B (Ref. 8) and Compactin 9) was isolated inde- pendently by two different groups from the cultures of Penicillium citrinum and Penicillium brevicom- pactum and proved to be a remark- ably potent HMG-CoA reductase inhibitor. Later, a closely related compound, lovastatin (Fig. 2,1b), formerly known as monacolin K (Refs 10, 11) and mevinolin 12 was isolated independently by two different groups from cultures of The atheromatous plaque or ather- oma is formed in part from lipid deposits, primarily cholesteryl esters, in the inner walls of arteries. Growth of the atheroma leads to constriction of the arterial lumen and ultimately results in atherosclerosis and coronary heart disease (CHD), the leading cause of death and disability in Western countries. Epidemiological evidence 1-3 strongly indicates that hyper- cholesterolemia - or more accur- ately, elevated plasma levels of low-density lipoprotein chol- esterol (LDL-C) - is a major risk factor for the development of CHD. These observations have stimu- lated intensive efforts directed towards the development of thera- peutic agents for prevention and treatment of atherosclerosis based on the attenuation of plasma cholesterol levels (see Ref. 4). Results of the recently com- pleted Lipid Research Clinics Coronary Primary Prevention Trial (LRC-CPPT) 5'6 provide strong sup- port for the rationale underlying this approach. This trial clearly demonstrated that the reduction of LDL-C through dietary modifica- tion and treatment with the bile acid sequesterant colestyramine, either alone or in combination, diminished the incidence of CHD morbidity and mortality in hyper- cholesterolemic men at high risk for CHD. Nevertheless, these mea- Ta-]yh Lee is Senior Research Fellow in the Department of Medicinal Chemistry, Merck Sharp & Dohme Research Laboratories, West Point, PA 19486, USA. 1987, Elsevier Publications, Cambridge 0165 6147/87/$02.00 sures often fail to lower elevated plasma LDL-C levels to the desired extent, particularly in patients with familial hypercholesterol- emia. Because most cholesterol in the body is synthesized de novo, the control of endogenous chol- esterogenesis would be an attract- ive and potentially more effective way to lower plasma cholesterol levels. Cholesterol is synthesized from acetyl-CoA via a series of more than twenty enzymatic reactions. The major rate-limiting step in this pathway is regulated by the act- ivity of the enzyme 3-hydroxy-3- methylglutaryl-coenzyme A re- ductase (HMG-CoA reductase, EC1.1.1.34) 7, which catalyses the conversion of HMG-CoA to mev- alonic acid (Fig. 1). Therefore, for several decades, this enzyme has been considered to be a prime target for pharmacological inter- vention for the control of endo- genous cholesterogenesis. The Monacus ruber and Aspergillus terreus. The subsequent isolation of a number of related compounds has been reported (see Ref. 13). Mevastatin and lovastatin are clearly the most extensively in- vestigated members of this family of compounds. The inhibition of HMG-CoA reductase by mevastatin and lova- statin is reversible and competi- tive with respect to HMG-CoA, the natural substrate. The Ki values of the dihydroxy acid forms of meva- statin and lovastatin (Fig. 2, 2a and 2b, which are the biologically active forms of mevastatin and lovastatin) are 1 nM and 0.6 riM, respectively. Neither compound affects other enzymes involved in cholesterol biosynthesis. In mam- malian cells cultured in a medium containing LDL, the synthesis of other biologically important sub- stances such as ubiquinone, doli- chol and tRNA, which also are derived from mevalonate and re- Acetate J Multiple Steps CH3 HO~'~-COzH SCoA H MG -CoA f HMG-CoA Reductase NADPH Cholesterol 'I Multiple Steps CH3 HO~7"-CO2H k,, OH Mevalonic Acid Fig. 1. Biosynthetic pathway for cholesterol.
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`TIPS - November 1987 [Vol. 8] quired for cell growth, is not affected even when the activity of HMG-CoA reductase is severely suppressed (up to 98%) by meva- statin 14,15. These findings strongly suggest that mevastatin and lovastatin are specific inhibitors of HMG-CoA reductase. However, of greatest in- terest is the finding that these two natural products are highly effective hypocholesterolemic agents in i 13 several animal spec es and humans 16-19. Not surprisingly, after the disclosure of these initial find- ings, efforts directed toward the synthesis of mevastatin, lovastatin and related analogs were initiated in many laboratories (see Ref. 20). Totally synthetic analogs of mevastatin Before the discovery of lova- statin, an effort was initiated in our laboratories directed towards the development of totally synthetic HMG-CoA reductase inhibitors devoid of the structural com- plexities of mevastatin. A model synthetic analog of mevastatin was formulated based largely on the following considera- tions. The structural similarity be- tween the dihydroxy acid moiety of structure 2 and HMG-CoA or mevalonic acid is readily recog- nized. However, since the Km value for HMG-CoA, the natural substrate of the enzyme, is approxi- mately 10-5M (about 10 4 weaker than the Ki values of mevastatin and lovastatin), the substituted polyhydronaphthyl moieties in mevastatin 21 and lovastatin must also play an important role in the inhibition of the enzyme. Based on this line of reasoning and the known facts about the HMG-CoA reductase-catalysed re- duction of HMG-CoA to meval- onic acid, our early model of a synthetic analog of structure 2 is represented by the generalized structure 3. It consists of a di- hydroxy acid moiety similar to that of structure 2a and a lipophilic group linked by either a saturated or an unsaturated 2-carbon bridge. Thus, structure 4, which is the corresponding lactone form of structure 3, was the initial target• It should be noted that the stereo- chemical requirements at positions C-4 and C-6 of lactone 4 were not known at this point. Although the chiralities of two steric centers in the lactone ring of mevastatin (cf. 443 H o .o o -I-H20 --H20 a, R -- H (Mevastatin) b, R = OH 3 (Lovastatin) H HO~"CO2H O l"Y. OH HO~co2H A I x A = CH2CH 2 or CH = CH X -- Aromatics HO~O W ° A I x 3 HO~r'~O i cI HO ~,./~O A I X 6 HO~co2Na ~..o. HO!~O! R" I,,,,~f6 R20 H r ,~ ~CH 3 CH 3" : 7 (SRI-62320) 8 HO~T,,~ CO2Na '-.y..OH o ~:.~~ I~CH3 OH 3 ...... HOy"",,CO2N a ~,,.oa O : HO" ~ ~ 10 (CS-514 or Sq-31000) Fig. 2. Structures of mevastatin, Iovastatin and the corresponding ring-opened di- hydroxy acid forms, and other analogs.
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`a a~ TIPS - November 1987 [Vol. 8] TABLE I. In-vitro inhibitory activities of lactones 6 (Fig. 2) towards HMG-CoA reductase Code Optical activity A X Relative potency* • + ' ......... :+ + ::'++. " ..... +- ........... ,i a + t-CH=CH 0.16 CI C + CH2CH2 F-~-C.,O~-C+ 38 T CI • ' :~,+ +:, d + t'CH=CH c~ 100 CI i : • :: .+ = .: .+ . e + t-CH=CH ~'~'~1 ~ 289 CH~ CH~ CH3 • All compounds in this table were tested after being converted to the sodium salts of the corresponding dihydroxy acids. The relative potency of the test compound was determined by compadng its IC~ value with that of mevastatin, which was tested simultaneously and arbitrarily assigned a relative potency value of 100. structure la) had been determined earlier 9, their critical importance to intrinsic inhibitory activity was yet to be ascertained. This issue was quickly resolved by determin- ing that: (1) all activity resides in the trans-diastereomer 6a(+) (i.e. the cis-diastereomer 5(+) is in- active); and (2) only enantiomer 6a(+) is active 22. Similar results were observed subsequently in every compound series examined (see Table I), indicating that the chiralities of the two steric centers in structure 6 are critical in deter- mining intrinsic HMG-CoA re- ductase inhibitory activity. Later, the finding that compound 6d(+) • possessed the same chirality in the lactone ring as that present in mevastatin was determined by X- ray crystallography 23 and further supported this conclusion. However, the weak intrinsic inhibitory potency of structure 6a(+) needed to be optimized. Attachment of either an arylmeth- oxy group (benzyl ether series) 24 or an aryl moiety (biphenyl series) 2s at the 6-position of the 2,4- dichlorophenyl ring in compound 6a(+) dramatically enhanced potency. In the benzyl ether series, introduction of a 4-fluoro group on the benzyl moiety induces a remarkable increase in potency. Later, this substitution proved to be equally useful in other series. Another significant contribution towards the improvement of potency was observed when com- pound 6b(+) was hydrogenated to 6c(+). Resolution of 6c(+) yielded 6c(+), which had an inhibitory potency approaching that of meva- statin. Even more profound enhance- ment of potency was observed when suitable aryl groups were placed at the 6-position of the 2,4- dichlorophenyl ring in 6a(+). For example, a 1750-fold increase in potency attended the formal con- version of compound 6a(+) to 6d(+) (Table I). In view of the toxicity ascribed to chlorinated biphenyls, it was deemed desir- able to find a suitable replacement for the chloro groups in 6d(+). The methyl group was considered to be a favorable bioisosteric replace- ment for the chloro group because of their comparable sizes and lipophilicities. Furthermore, aromatic methyl groups are susceptible to bio- oxidation and the resultant met- abolite(s) is expected to be more readily eliminated from the body. Indeed, replacement of the chloro- substituents in structure 6d(+) with methyl groups and further refinement ultimately afforded compound 6e(+), which has an inhibitory potency almost three times that of mevastatin. It is also noteworthy that a substantial de- crease in potency occurs when structure 6e(+) is reduced to 6f(+). This result is contrary to earlier observations in the benzyl ether series and the prediction of an advantage in having a saturated or an unsaturated 2-carbon bridge connecting the lactone and the lipophilic groups remains pre- carious. Finally, the recent dis- closure of research on compounds related to compound 7 (SRI- 62320) 2s appears to be particularly interesting. As evidence of the therapeutic usefulness of HMG- CoA reductase inhibitors con- tinues to mount, it is anticipated that research activities in search of new and novel inhibitors will be greatly intensified in the future. TABLE II. In-vitro inhibitory activities of lactones 8 (Fig. 2) towards HMG-CoA reductase Code R ~ R 2 Relative potency* O b H CH,CH,C-~- 254 H CH~ ....... ® d H F.~-c~ - 119 +++, +++, *See Table I.
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`TIPS - November 1987 [Vol. 8] 445 Reductase inhibitor Bile acid HMG CoA + Untreated depletion Reductase inhibitor bile acid depletion Plasma Liver Intestine (, ~ U Fig. 3. Rationale for the therapeutic treatment of hypercholesterolemia. Semisynthetic analogs of lovastatin Lovastatin (lb) 12 differs from mevastatin (la) structurally only by the presence of a 60c-methyl group and its relative potency is approximately twice that of meva- statin. Its discovery added a new direction to research in this area- delineation of the structural fea- tures of lovastatin responsible for its remarkable inhibitory activity and the use of this information to develop even more potent and efficacious inhibitors. Studies on the modification of the 2(s)-methylbutyryl moiety of lovastatin 26 yielded the following conclusions: • The presence of a side chain ester moiety is crucial to in- hibitory potency (the potency of compound 8a is very weak). • The stereochemistry at the car- bon 0c to the ester carbonyl group is not critical, since diastereomer 8b has the same potency as that of lovastatin (lb). • The spatial requirements of the acyl moiety are compatible with compact, branched aliphatic acyl groups. • Additional branching at the 0c- carbon of the acyl moiety en- hances potency. In particular, the introduction of an addi- tional methyl group on the carbon 0c to the ester carbonyl in lovastatin afforded structure 8c (MK-733) which, with an intrin- sic potency more than six times that of mevastatin, is the most potent HMG-CoA reductase in- hibitor reported to date. A series of side chain ether analogs of lovastatin were found to be mostly weaker inhibitors 27, although compound 8d, the best in the series, shows a respectable level of potency. Introduction of a methyl group (0~-orientation) to the hydroxy-bearing carbon of the lactone in lovastatin provides homolog 8e (Ref. 27) in which the lactone moiety bears a close struc- tural resemblance to HMG-CoA and mevalonic acid, the respective substrate and product of the reduction catalysed by HMG-CoA reductase (Fig. 1). Surprisingly, this modification is detrimental to activity. This was also true when similar modifications were made in the synthetic analogs. Insertion of a methylene unit between the lactone hydroxy-bearing carbon and the carbonyl group in lova- statin afforded the carboxylate 9, which was devoid of intrinsic HMG-CoA reductase inhibitory activity. Finally, minimal decreases in potency were observed when either one or two double bonds in the hexalin moiety of lovastatin were hydrogenated, providing that the ring juncture of the resultant hydrogenation product is trans; when the ring juncture is cis, activity is markedly diminished. A most noteworthy finding has been the recent discovery of CS-514 (structure 10, also known as SQ- 31000) 28, a microbial oxidation product of mevastatin. CS-514 is currently undergoing clinical evaluation. The initial reports from investigations of this new HMG- CoA reductase inhibitor show promise. Therapeutic potential as hypocholesterolemic drugs One of the major recent develop- ments in the prevention and treatment of CHD is the under- standing of how plasma chol- esterol levels, especially LDL-C levels, are controlled. Based on the findings of M. S. Brown and J. L. Goldstein (see Ref. 29), a rationale for the therapeutic treatment of hypercholesterolemia is schem- atically presented in Fig. 3. A liver cell normally receives its chol- esterol from: (1) uptake from circulating LDL-C via receptor- mediated endocytosis; (2) de-novo biosynthesis; and (3) uptake from chylomicron remnants. It then converts much of its cholesterol pool into bile acids. A large fraction of the bile acids secreted from the liver is returned to the liver through enterohepatic cyc- ling. It is well established that the liver is the major site of LDL receptors and the production of LDL receptors is driven by the liver cell's demand for cholesterol. An effective therapeutic approach for lowering plasma LDL-C levels should be focused on ways to increase the production of LDL receptors in the liver. This is achievable by inhibition of the intestinal reabsorption of bile acids and/or inhibition of en- dogeneous cholesterol synthesis. Ultimately, when a new steady state is attained, the absolute amount of cholesterol entering the liver through the receptor pathway remains approximately the same as before, because the fall in plasma LDL levels is balanced by the increase in LDL receptors. But the important difference is that this delivery is now occurring at a lower plasma LDL level. In practice, the ingestion of bile acid resins (e.g. colestyramine and colestipol) causes plasma LDL
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`446 TIPS - November 1987 [Vol. 8] levels to decrease by blocking the reabsorption of bile acids and thereby increasing the metabolism of cholesterol. However, the effect of this therapy is not profound. It usually produces only about a 20% drop in plasma LDL-C levels 5"6. The second method, using HMG- CoA reductase inhibitors such as mevastatin 16 and lovastatin 17-19 to inhibit cholesterol synthesis, is a much more effective approach for lowering plasma LDL levels. When given as a single agent (40 mg per day) to familial hyperchol- esterolemic heterozygotes, lova- statin decreases plasma LDL levels by 33% (from 361 + 13 mg cm -3 to 242 + 15 mg cm-3) 19. When given together with colestipol, an even greater drop (46%) in plasma LDL levels was reported 19. The hypo- cholesterolemic effect of lovastatin has been ascribed to the increased production of LDL receptors re- suiting from the inhibition of de-novo cholesterogenesis. Other studies also suggest that the synthesis of very low density lipoprotein (VLDL), a precursor of LDL, has also been suppressed by lovastatin (D. R. Illingworth, pers. commun.). Recently, lova- statin was also found to be effec- tive in the treatment of patients with nonfamilial hypercholester- olemia 3°. Specifically, patients re- ceiving 40 mg twice a day experi- enced a mean reduction in plasma LDL levels of 40%. So far, consis- tently good patient acceptance and a low incidence of side effects have been observed in all studies with lovastatin (for detailed discussion, see Ref. 31). The important prin- ciple that has been realized from these studies is that stimulation of LDL receptor activity ultimately lowers plasma LDL-C levels with- out grossly altering cholesterol supply to cells. HMG-CoA reduc- tase inhibitors are powerful means of taking advantage of this prin- ciple in the effective reduction of plasma LDL levels. [] [] [] Evidence relating plasma chol- esterol levels to atherosclerosis and CHD has become compelling. Mechanisms responsible for high concentrations of LDL are becom- ing understood. The LDL receptor is recognized as an important ele- ment in the regulation of plasma LDL levels. The fact that HMG- CoA reductase inhibitors are highly effective hypocholesterol- emic agents is now well estab- lished. The use of this newly dis- covered therapy to treat a fraction of the population at high risk of CHD because of extremely high plasma LDL levels (e.g. familial hypercholesterolemia patients) appears to be widely accepted by the medical community. Will HMG-CoA reductase inhibitors also become the choice of therapies to treat moderate hyperchol- esterolemia, which is a much more common health problem than familial hypercholesterolemia? In- tensive efforts to establish the long-term clinical safety of HMG- CoA reductase inhibitors are clearly vital to this issue. References 1 Kannel, W. B., Castelli, W. D., Gordon, T. and McNamara, P.M. (1971) Ann. Intern. Med. 74, 1-12 2 Keys, A. (1980) in Seven Countries: A Multivariate Analysis of Death and Coronary Heart Disease, pp. 1--381, Harvard University Press 3 Stamler, J., Wentworth, D. and Neaton, J. D. (1986) J. Am. Med. Assoc. 256, 2823- 2828 4 Prugh, J. D., Rooney, C~ S. and Smith, R.L. (1983) Annu. Rev. Med. Chem. 18, 161-170 5 LRC-CPPT (1984) J. Am. Med. Assoc. 251, 351-364 6 LRC-CPPT (1984) J. Am. Med. Assoc. 251, 365-374 7 Rodwell, V. W., Nordstrom, J. L. and Mitschelen, J. J. (1976) Adv. Lipid Res. 14, 1-74 8 Endo, A., Kuroda, M. and Tsujita, Y. (1976) J. Antibiot. 29, 1346-1348 9 Brown, A. G., Smale, T. C., King, T. J., Hasenkam, P. and Thompson, R.H. (1976) J. Chem. Soc. Perkin Trans. 1,1165- 1170 10 Endo, A. (1979) J. Antibiot. 32, 852-854 11 Endo, A. (1980) J. Antibiot. 33, 334-336 12 Alberts, A. W., Chen, J., Kuron, G., Hunt, V., Huff, J., Hoffman, C., Rothrock, J., Lopez, J., Joshua, H., Harris, E., Pachett, A., Monaghan, R., Currie, S., Stapley, E., Albers-Schonberg, G., Hen- sen, O., Hirshfield, J., Hoogsteen, K., Liesch, J. and Springer, J. (1980) Proc. Natl Acad. Sci. USA 77, 3957--3961 13 Endo, A. (1985) J. Med. Chem. 28, 401-405 14 Faust, J. R., Brown, M. S. and Go!dstein, J. L. (1980) J. Biol. Chem. 255, 6546-6548 15 Brown, M. S. and Goldstein, J. L. (1980) J. Lipid Res. 21, 505-517 16 Mabuchi, H., Sakai, T., Sakai, Y., Yoshi- mura, A., Watanabe, A., Wakasugi, T., Koizumi,. J. and Tekeda, R. (1983) N. Engl. J. Med. 308, 609-613 17 Bilheimer, D. W., Grundy, S. M., Brown, M.S. and Goldstein, J.L. (1983) Proc. Natl Acad. Sci. USA 80, 4124-4128 18 lUingworth, D. R. and Sexton, G. J. (1984) J. Clin. Invest. 74, 1972-1978 19 Illingworth, D. R. (1984) Ann. Intern. Med. 101, 598-604 20 Rosen, T. and Heathcock, C. H. (1986) Tetrahedron 42, 4909---4951 21 Nakamura, C. E. and Abeles, R. H. (1985) Biochemistry 24, 1364-1376 22 Stokker, G. E., Hoffman, W. F., Alberts, A.W., Cragoe, Jr, E.J., Deana, A.A., Gilfillar~, J. L., Huff, J. W., NoveUo, F. C., Prug~, J. D., Smith, R. L. and Willard, A. I~:,(1985) J. Med. Chem. 28, 347-358 23 Stokl~er, G. E., Alberts, A. W., Anderson, P.S., Cragoe, Jr, E.J., Deana, A.A., Gilfillan, J. L., Hirshfield, J., Holtz, W. J., Hoffman, W. F., Huff, J. W., Lee, T.-J., Novello, F.C., Prugh, J.D., Rooney, C.S., Smith, R.L. and Willard, A.K. (1986) J. Med. Chem. 29, 170-181 24 Hoffman, W. F., Alberts, A. W., Cragoe, Jr, E.J., Deana, A.A., Evans, B.E., Gilfillan, J. L., Gould, N. P., Huff, J. W., Novello, F. C., Prugh, J. D., Rittle, K. E., Smith, R. L., Stokker, G. E. and WiUard, A. K. (1986) J. Med. Chem. 29, 159-169 25 Engstrom, R. G., Weinstein, D. B., Kathawala, F. G., Scallen, T., Eskesen, J. B., Rucker, M. L. and Miserendino, R. (1986) IX International Symposium on Drugs Affecting Lipid Metabolism, Florence, Italy, p. 26 (Abstr.) 26 Hoffman, W. F., Alberts, A. W., Anderson, P. S., Chen, J. S., Smith, R. L. and WiUard, A. K. (1986) J. Med. Chem. 29, 849-852 27 Lee, T.-J., Holtz, W. J. and Smith, R. L. (1982) J. Org. Chem. 47, 4750--4757 28 Tsujita, Y., Kuroda, M., Shimada, Y., Tanzawa, K., Arai, M., Kaneko, I., Tanaka, M., Masuda, H., Tarumi, C., Watanabe, Y. and Fujii, S. (1986) Bio- chim. Biophys. Acta 877, 50-60 29 Brown, M. S. and Goldstein, J. L. (1986) Science 232, 34-47 30 Lovastatin Study Group II (1986) J. Am. Med. Assoc. 256, 2829-2834 31 Tobert, J. A. (1987) Circulation 76, 534- 538 TiPS centrefolds Full colour reprints of the following centrefolds published earlier in T/PS are available: Dopamine receptor subtypes Eicosanoids (updated May I987) Functional receptors for 5-HT Peptides, receptors, antagonists Price per 5 copies is £6.00 (+ 15% VAT in UK) or US $9.00 (incl. p. & p.). Larger orders (100+) available at 15% discount. Orders from: Elsevier Publications Cambridge, 68 Hills Road, Cambridge CB2 1LA, UK.
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