CA2257084A1 - Treatment of lipid and glucose metabolism disorders with dopamine and serotonin agonists - Google Patents

Abstract

The present invention relates to novel, improved methods for regulating in a mammal at least one of body weight, body fat, food consumption, lipid metabolism and glucose metabolism. Administration of a D1 or D2 agonist or both in the morning, combined with administration of a 5HT1B receptor subtype agonist at night results in reduction in food consumption, body weight, serum glucose levels and total body fat.

Description

TREATMENT OF LIPID AND GLUCOSE METABOLISM DISORDERS
WITH DOPAMINE AND SEROTONIN AGONISTS

FIELD OF THE INVENTION
This invention relates to novel, improved methods for modifying or 10 regulating in a subject (vertebrate animal or human) at least one of body weight, body fat, food consu~lplion, lipid metabo}ism, and glucose metabolism.

BACKGROUND OF T~E INVENTION
Obesity and Lipid Metabolism Di~. de.~ - Body Fat Loss In hllm~n~, obesity can be defined as a body weight e~cee~ling 20% of 15 the desirable body weight for individuals of the same sex, height and frame (Salans, L.B., in Endocrinolo~y & Metabolism, 2d Ed., McGraw-Hill, New York 1987, pp.
1203-1244; see also, R.H. Williams, Textbook of Endocrinolo~y, 1974, pp. 904-916).
In other Anim~lc (or also in hllm~n~) obesity can be determined by body weight p~tt~rns correlated with prolactin profiles given that members of a species that are 20 young, lean, and "healthy" (i.e., free of any disorders, not just metabolic disorders) have daily plasma prolactin level profiles that follow a pattern ch~racteri~ti~ of the species. This pattern is highly reproducible with a small standard deviation. Members of a species surrel~ g from lipid and/or metabolism disorders, however, have aberrant prolactin profiles that depart from the normal (or healthy subjects') pattern by at least 25 1 SEM in at least two spaced apart time points or by at least 2 SE~ (standard error of the mean) in at least one time point.
Obesity, or excess fat deposits, correlate with and may trigger the onset of various lipid and/or glucose metabolism disorders, e.g. hypertension, Type IIdiabetes, atherosclerosis"~lin~allly etc.

Wo 97/46239 PcTruss7llo838 Even in the qhsenl~e of clinical obesity (according to the above dPfinition) the reduction of body fat stores (notably visceral fat stores) in man, ecreciqlly on a long-term or p~" ~ n~.nt basis would be of cignific~nt benefit, both cosnletievlly, physiologically and psychologically.
The reduction of body fat stores in domestic animals (as well as pets), e~eciq-lly on a long-term or 1~ qnP~lt basis, would also obviously be of conci~er~h economic benefit to man, particularly since farm animals supply a major portion of man's diet; and the animal fat may end up as de novo fat deposits in man.
Whereas controlled diet and exercise can produce modest results in the reduction of body fat ~lepocit~, prior to the cl~mlllqtive work of the present inventors (inclur~in~ the prior co-pending patent applic~tion~ and issued U.S. patents lerell~ to below), no truly effective or practical treatment had been found for controlling obesity or other lipid metabolism disorders.
Hyperlipoproteine~ is a condition in which the concentration of one or more of cholesterol- or triglyceride-carrying li~r~leills (such as chylomicrons, very low density lipopluteins ("VLDL"), and low-density lipupluteihls ("LDL") in plasma ~cee~lc a nor nal limit. This upper limit is generally defined as the ninety-fLfth lut;r~e~llile of a random population. Elevated levels of these substances have also been positively correlated with atherosclerosis and the often resulting cardiac infarction, or "heaIt attack", which accounts for a~lv~ at~ly half of all deaths in the United States. Strong clinical evidence has been pr~senled which correlates a re~luction in plasma ~uoploleill conc~ntrqti()n with a reduced risk of atherosclerosis (Noma, A., et al., Atherosclerosis 49:1, 1983; Illingworth, D. and Conner, W., in Endocrinology &
Metabolism, McGraw-Hill, New York 1987). Thus, a ~ignifi~qnt amount of ~s~;h 25 has been devoted to finding L~lP-~I methods which reduce levels of plasma choles-terol and triglycerides. High LDL and/or VLDL accompqni~d by high triglyceride levels in the blood con~lilule most iulpolL~u~L risk factors for atherosclerosis.
Re-luctil~n of one or both of li~uu~o~eih~s and triglycerides in the blood would reduce the risk of atherosclerosis and cardiac arrest, or retard their development.
Another subset of the plasma li,uo~rule~ns found in vt~ al~,i, are high density lipoploleills, HDL ("HDL"). HDL serve to remove free cholesterol from the plasma. A high HDL co-~c~.l.,.l;on, as a ,u~ ;elllage of total plasma cholesterol, has been qcso~ ~ with a reduced risk of all~e~sclerosis and he~t disease. Thus, HDL
are known in the lay press as "good" chol~sterol. The~ful~, t~ GI~IiC ~Llàl~.gies involve atle~-pls both to reduce plasma LDL and VLDL content (that is, reduce total plasma chole~stçrol), and to increase the HDL fraction of total plasma cholesterol.
Several lines of research in(~ te that simply increasing HDL is of benefit even in the absence of LDL or VLDL reduction: Bell, G.P. et al., Atherosclerosis 36:47-54, 1980; Fears, R., Biochem. Pharmacol 33:219-228, 1984; Thol.")son, G., Br. Heart J.
51:585-588, 1989; R1~khurn, H. N.E.J.M. 309:426-428, 1983.
Current therapies for llyl~e~ ul~lote;~ include a low fat diet and elimin~tic)n of aggravating factors such as sedent~ry lifestyle. If the hy~;llipoprole;nPnni~ is secondary (i.e., inri~ent to e.g., a d~ficien~y of lip~pr~
10 lipase or LDL receptor, various endocrine pathologies, alcoholism, renal disorders, or hepatic disorders) then control of the underlying disease is also central to L~ .e~
HyperlipoproleillPnni~s are also treated with drugs, which usually alter the levels of particular components of the total plasma cholestProl, as well as reduce the total plasma lipid component. Among the most recently introduced drugs to treat 15 hyperlipoplvlei-~P~ is lovastatin (MEVACOR~9) which selectively inhibits an enzyme involved in cholesterol production, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reduct~ce. This drug specifically reduces total cholesterol and can cause amodest (5-10%) increase in HDL concenllations~ However, benefits from these therapies vary from subject to subject~
Moreover, use of the HMG-CoA enzyme inhibitor is sometimes acco",r~ d by side effects such as liver toxicity, renal myoglobinuria, renal shutdown, and lPnti~ul~r opacity. The risk of such side effects nPces~ es close monitoring of the patipntc (e.g., liver function is tested monthly).
Another drug prescribed against hyperlipopn~lçi~ is clofibrate. The 25 effectiveness of clofibrate also varies from subject to subject and its use is often accol"~-i~ by such side effects as neph~lic syndromes, myalgia, nausea, and abdominal pain.

Type II D;&bltes (NIDDM) and Glucose Metabolism Disorders Type II (li~betes, also known as non-insulin dependent diabetes mellihls 30 (NIDDM), is one of the most insidious of the major ~ e~ces. NIDDM can strike s~lddenly or lie lln~ nQsed for years while ~tt~1ring the blood vessels and nerves.
Individuals ~urrt;~ g from NIDDM, as a group, are far more often ~fflirtP,d with CA 022~7084 1998-12-02 blin-lness, heart disease, stroke, kidney disease, h~ring loss, gangrene, and impotence. One third of all visits to physicians are occasioned by this disease and its complications, and diabetes and its com~ ti~ n.~ are a leading cause of l~ntimely death in the United States and in the Western world.
S NIDDM adversely affects the way the body uses sugars and s~hes which, during ~ligestit n, are converted into glll~ose. Tn~-llin, a honnone produced by the pancreas, makes the glucose available to the body's cells for energy. In muscle, adipose (fat), and comlecLive tissues, insulin f~cilit~tes the entry of glucose into the cells by an action on the cell membranes. The ingested glucose is normally converted 10 in the Iiver to CO2 and H2O (50%); to glycogen (5%); and to fat (30-40%), the latter being stored in fat deposits. Fatty acids from the adipose tissues are circulated by the bloodstream, returned to the liver for re-synthesis of triacylglycerol, and metaboli~ed to ketone bodies for utilization by the tissues. The fatty acids are also metabolized by other organs. Fat formation is a major pathway for carbohydrate utili7~tion.
The net effect of insulin is to promote the storage and use of carbohydrates, protein, and fat. Insulin deficiency is a common and serious pathologic condition in man. In insulin-dependent diabetes (IDDM or Type I diabetes) the pancreas produces little or no insulin, and insulin must be injected daily for the survival of the diabetic. In noninclllin-dependent diabetes the pancreas retains the 20 ability to produce insulin and, in fact, may produce higher than normal amounts of insulin, but the amount of insulin is relatively insufficient, or less than fully effective, due to cellular reCict~nce to insulin, i.e. cells do not respond to normal levels of insulin by taking up glucose, amino acids, and fat.
In either form of diabetes, there are widespread abnorrn~lities. In most 25 NIDDM subjects, the fun~mental defects to which the abnonn~lities can be traced are (I) a reduced entry of glucose into various "yeli~he,dl" tissues and (2) an increased liberation of glucose into the circulation from the liver. Thcl~rcl~, there is an extracellular glucose excess and an intracellular glucose deficiency. There is also a decrease in the entry of amino acids into muscle and an increase in lipolysis.
30 Hyy~lliyoproteinemia is also a complication of di~betes. The cum~ tive effect of these NIDDM-associated abnorm~liti~s over time is severe blood vessel and nerve damage which can lead to blindn~s~ and gangrene, among other afflictions.

Other than the m.o.tht~1c of the present invention and previous work by the present inventors (~liccucc-e~ below), no effective tre~tm~nt has been found for controlling either hy~ .linP.mi~ or insulin recistAnce. Hyye~ ..linemi~ is a higher-than-normal level of insulin in the blood. Insulin les;~ e can be defined as a state S in which a normal amount of insulin produces a sllknnrm~l biologic response by cells.
In insulin-treated patiPntC with ~ 7et~s, insulin rÇcist~nce is con~i~lered to be present whenever the tk~,...peu~ dose of insulin e ~-~eeds the secretory rate of insulin in normal persons. Insulin re~;ct~nre is also associated with higher-than-normal levels of insulin i.e. hyperin.c~llin~mi~ -- when normal or elevated levels of blood glucose are present.

~viu~s Work in This Field Studies by the present inventors and others have in~ t~l that the naturally occurring annual cycle of body fat stores, pervasive among ve,lel,l~tes in the wild, reflects the activities of an adjustable central metabolistat that is comprised of circadian hypoth~l~mic neural components, among which are dopanlille.~ic and 15 serotonergic activities. Changes in the phase-relationships of these circadian do~ rgic and sel~toner~ic activities induce seasonal changes in metabolism.
These circadian activities can be adjusted by a~ iately timed tre~tm. ntc with hormones or neurotrAncmitt~r-affecting drugs. In this regard, timed A~lminictrAtiQn of bromocriptine, a sympatholytic dopalllh~e D2 receptor agonist ("D2 agonist") with a!2 20 adrenergic receptor agonist ("lY2 agonist") and ~, adrenergic receptor antagonist ("(x antagonist") activities, as well as serotonin ("S-hydroxyll~y~lline" or "SHT") inhibiting activities, has been demonstrated to reduce body fat stores in a variety of Anim~lc including hllm~nc without a concomitant reduction in food col~cnmptiQn.
Timed a~1ministration of bromocriptine has also been found to reduce llyyc.;i-c~11inemi~, 25 hy~e~ idemia, and glucose intolerance.
The present inventors and their co-workers have previously discovered that timed a~lminictration (i.e., ~-lmini~tr~tion at a time of day when the body, because of circadian neurooscillations, is most responsive to a bioactive agent) of either or both of (i) certain prolactin re~ cing dopamine (D2) agonists such as bromocriptine and (ii) prolactin-increasing substances such as CA 02257084 l998-l2-02 (a) dopalllu~e antagonists, such as metoclopr~mi~le and (b) sel(Jtonill agonists and precursors, in~ rling by way of non-iling e~r~mpl~ 5-hydl~ylly~lo~han reduce body fat stores, obesity, plasma triglycerides, and cholesterol as well as 5 hy~ lyce~nia, hy~ linP.mi~, and insulin le3;~ e. This work is disclosed in U.S. Patent Nos. 4,659,715; 4,749,709; 4,783,469; 5,006,526.
It is pr~r~ led to a~lminict~r the prolactin red~lçing s~b~l~nce~ at a first pred~ i..ed time of day to effect a decrease in the circul~ting prolactin levels of the subject to be treated during an interval within the subject's daily prolactin cycle or rhythm when circulating (blood) prolactin levels are low in young, healthy subjects of the same species, thereby causing the prolactin rhythm of the treated subject toapproach or to conform to the standard or healthy subjects' prolactin rhythm. It is also preferred to ~rlminict~r the prolactin-increasing substances at a second prede~ ed time of day to effect an increase in the circul:lting prolactin levels of the subject to be treated during an interval within the subject's daily prolactin cycle or rhythm when circ~ ting (blood) prolactin levels are high in young healthy subjects of the same species, thereby causing the prolactin rhythm of the treated subject to approach, or conro~.ll to, the standard or healthy subjects' prolactin rhythm. Such methods are disclosed in U.S. Patent Nos. 5,468,755; 5,496,803; 5,344,832; 5,585,347, USSN
08/456,952, and PCT applications US93/12701 and US95/09061.
It is also known in the art that some of the effects of bromocriptine are supported by endogenous dopamine. ~Ergot Compounds and Brain Functions Neu.ullsychiatric Aspects: Advances in Biochemical Psychopharmacolo~y. M.
Gol~tein et al., Eds. (Raven Press, New York, 1980) vol. 23). Specifically, it has been shown that locomotor s~im~ tion and stereotyped behavior responses to bromocriptine are blocked by depletion of endogenous dopalllil~e in rodents. However, if a Dl dop~,~ihle receptor agonist is subsequçntly providçd to dopamine depleted ~nim~l~, the responsiveness to bromocri~tme is restored. Jackson, D.M. et al., Psychopharmacolo~y 94:321 (1988)). A similar dopaminergic D2:D, interaction has been demonstrated in dopaminergic inhihition of feeding behavior. Although thesestudies CO~ lll the ill~o~ ce of a D2:DI interaction in the activation of dop~minPrgic activities, the increased locomotor activity and decreased feeding response to D2:D, agonists is acute and short lived, lasting for only a few hours. (Cooper, S.J. et al., in W O 97/46239 PCT~US97110838 Dl:D2 Dul~allline Receptor Interactions, J. Waddington, Ed. (~c-q~emic Press, London, 1993) pp. 203-234).
Previûus work by third parties with D, and D2 dopamine agonists in combination has not demon~t~ted any effects on lipid and glucose metabolism, and has not produced long-term l~onses of d~ gic activities. The present inventors discovered that the conjoined admini~t~ion of a Dl dopamine agonist and a D2 d~lline agonist (or at least one of an cYl antagonist, an a!2 agonist and a s~olollel~ic inhibitor) in the morning results in an improvement in one or more of the metabûlic indices related to lipid and glucose metabolism when co-llp~d to the improvement (if any) provided by ~11mini~tration of a dopamine D2 agonist, such as bromocriptine, . d alone.
The present inventors have now ~InPxrectelly discovered that ~Imini~tration of a Dl or D2 agonist (or both) in the morning (e.g., between the hours of 5:00 and 13:00), combined with q.-~mini~tr.q-tion of a SHTIB receptor subtype agonist ("5HTlB agonist") at night (e.g., between the hours of 17:00 and 0:00) results in a surprising and unexpected reduction in food consumption and body weight. Such q~lminictr.q,ti(ln also results in a decrease in one or more metabolic indices related to lipid and glucose metabolism, such as serum glucose levels, total body fat, and body weight.
The present invention, which utilizes specific SHTIB receptor subtype agonists, provides ~ignifi~qnt advantages over the a~mini~tration of 5-HTP to patients.
Since the SHTlB agonists act specifi~?lly at the SHT,B receptor subtype, side effects associated with a nonspecific increase in selolo..il~ associated with non-specific 5-HTP
, ~mini~tr.qtion can be avoided.
OBJECTS OF THE INVENTION
It is an object of this invention to provide adrlitiQnql improved methods ~ for re~uçing in ve~ dt~ subjects (inchltling hllmqn~) in need of such trP-q~tmPnt at least one of food consumptiûn, body weight, body fat, plasma or blood glucose, and blood insulin.
Another object of this invention is to provide methods for reclucin~ at least one of insulin resictqnce (illlp~ed glucose tolerance), hyperinculinPmiq and , W O 97/46239 PCTrUS97110838 hy~e~lycell~ia, and glycosylated hemoglobin (in~ ling AlC), and abating Type II
etes or Syndrome X.
A further object is to provide methods for redll~ing or re~.li,~g or a~-csLi"g atherosclerosis by red~l-ing at least one of hy~ )oplulçi~emi~ and elevated blood triglycerides and/or cholesterol.
It is another object of this invention to provide methods for modifying and regulating lipid and glucose metabolism in a Jllanne blqnPfi~i~l to the subject.
It is still another object of the invention to provide methods for modifying and regulating lipid and glucose metabolism to provide effective tre~tm~ t~
for obesity or weight reduction.

SUMMARY OF THE INVENTION
It has now been found that at least one of the foregoing objects can be accomplished by (A) ~l1mini~tçring~ in the morning, to a subject in need of such treatment at least one of the following:
(i) a dopamine Dl agonist or a dopamine Dl agonist in conjunction with one agent or agent combination selected from groups (ii) through (iv);
(ii) a dopa,lline D2 agonist;
(iii) at least one of an adltnelgic al antagonist, an adlene gic cY2 agonist, and a serotonergic inhibitor;
(iv) a dopanline D2 agonist further conjûined with at least one of an adrenergic ~, antagonist, an adrenergic a2 agonist, and a selvlonelgic inhibitor; and also (B) ~1minictPring to the subject a SHTIB agonist.
Preferably, the fo~oing agents in (i), (ii), (iii) or (iv) above ("conjoined agents") are arlmini.~tered at a predet~.rminP~ time of day, i.e., within a restricted portion of a 24-hour period, preferably in the morning (i.e. close to the time of light onset). Since the dopa"l"le D, agonist amplifies the effect of the other agent or agents, the Dl agonist is also preferably ~dministered at about the same time. The SHTIB agonist should be a~1minict~red at night.

CA 022F,7084 1998-12-02 The ~ .h~ on of a dopamine Dl or D2 agonist in the morning and the ~minict~ti~ n of a SHT,B agonist at night resu1ts in s.lb,l~ 11y ~ugmPntP~ and in fact often synergistic, effects in improvement of one or more metabolic indices related to glucose or lipid metaboIism, and thus an improved modification or regulation of at least one of glucose metabolism, lipid metabolism, food consumption, body fat orweight gain.
Where a D2 agonist is employed, it is preferably an ergot ~lk~lo;d~ most .~feldbly bromocAptine.
In another aspect, the present invention is directed to a~lminictering to a 10 subject in the morning a D2 agonist and a SHTIB agonist at night, preferably at or shortly before a subject's bedtime.
It has been found that the additional ~r~minictration of a SHTIB agonist at night can effect a greater improvement on one or more of the fo~going metabolic indices than arlministr~tion of only a Dl and/or D2 agonist in the mon~ing.

Figure l depicts body weight gain or loss in leptin-deficient mice as a result of tr~P~tmpnt with various Dl, D2, and SHT agonists, and combinations thereof.
Figure 2 depicts fat body mass in leptin-deficient mice as a result of tr~P~tm~P,nt with various Dl, D2, and 5HT agonists and combinations thereof.
Figure 3 shows lean body mass in leptin-deficient mice as a result of tre~tmPnt with various Dl, D2, and 5HT agonists and combinations thereof.
Figure 4 shows the effect on food consu,l,l)lion over the test period in leptin-deficient mice as a result of tre~tment with various Dl, D2, and SHT agonists and combinations thereof.
Figure 5 is a depiction of blood glucose at the end of treatment in leptin-d~PficiPnt mice as a result of tre~tment with various Dl, D2, and 5HT agonists, and comhin~fiQns thereof.
Figure 6 depicts serum free fatty acids at the end of the treatment period in leptin-d~PficiPnt mice as a result of tre~ment with various Dl, D2, and SHT agonists, 30 and combin~tion~ thereof.

Figure 7 is a plot of body weight in rats on the first and last day of tre~tmP.nt with a vehicle control, a D2 agonist with a D, agonist, a D2 agonist with a SHT~B agonist, and a Dl agonist with a 5HT,B agonist. Vehicle at 1 and 11 HALO
(hours after light onset), BC at 1 HALO plus SKF 38393 at 11 HALO, BC at 1 HALO
plus SKF77434 at 11 HALO, or SKF 38393 at 1 HALO plus RU24969 at llHALO by injection on body weight in rats during nine days of daily tre~tm~nt at 1 and 11 hours after light onset.
Figure 8 is a plot of food consumption, in grams, of the rats treated as described in Figure 7 over the tre~tment period.

All litGl~tulG and patents and patent applications cited herein are incorporated by lGrGlGnce in their enlih~Gly. In case of a conflict, the present disclosure including its definitions shall control.
In one embodiment of the method of the present invention, a Dl 15 dopamine agonist is ~flmini~tered, preferably in the morning, and a SHT,B agonist is ~1mini~t~red at night, to a subject in need of tre~tm~nt In another embodiment of the method of the present invention, a D2 dopamine agonist is ~lmini~tered, preferably in the morning, and a SHTIB agonist is ~lmini~tered at night, to a subject in need of treatment.
In yet another embodiment of the method of the present invention, a Dl dopamine agonist is a~lmini.st~red in conjunction with a D2 dopa,lline agonist, preferably both are ~rlmini.stered in the morning, and a SHTIB agonist is a~mini~tered at night, to a subject in need of In a further embodiment of the method of the present invention, a Dl 25 dopamine agonist is ~lmini.ctered in conjunction with a second agent, con~i~ting of at least one of a D2 agonist, an (X2 agonist, an ~1 antagonist, and a serotonergic inhibitor (or a D2 agonist and at least one of the rem~ining agents) preferably in the morning, and a SHTIB agonist is ~rlmini.~tered at night, to a subject in need of tre~tmPnt As used herein and applied to allmini~tration of more than one active 30 ingredient the terms "conjoined" or "in conjunction" mean that the subject being thus treated receives a first active agent and also at least one other active agent, but not n.o~ess~rily within the same fonnnl~ti-)n or dosage form and not neces.c~rily at the same .J~ ;s~ n time. For el~mple, the D, agonist and D2 agonist or the other agent(s)can be ~dmini~tered at the same time (in the same dosage form or in two or more divided dosage forms) or sequ~nti~lly at dir~e enl times and in dil~r~lelll dosage forms.
Two subtypes of dopamine ~ce~lol~, D, like and D2 like, were S idPntifie~ on the basis of ph~rrn~cological and bioch~.mi-~l criteria (see, e.g., Kebabian and Cave, Nature, 277:93 (1979)). What is meant by a "Dl do~l,ine agonist" or "D, agonist" is a compound which is capable of activating or pol~ g D, do~alnine receptors (e.g. Dl-like receptors such as Dl and D5 dopamine l~eplor~). See, e.g., Deany et al., Nature 347:72-76 (1990), Monsma, et al. Proc. Natl. Acad. Sci. USA87:6723-6727 (1990), Sunahara, et al., Nature 347:80 (1990), Zhou, et al., Nature 347:76-80 (1990).
Suitable assays for testing compounds for D, or D2 agonist activity (e.g., receptor binding assays asnd biological functional assays) are well known in the art, see. e.g., Javitch, et al., Proc. Natl. Acad. Sci. USA 91:10355 (1994); Piercey, et al., Eur. J. Pharm. 317:29 (1996); Paveney, et al., Eur. J. Pharm., 317:175 (1996);
Lovenberg, et al., Eur. J. Pharm., 166:111 (1989); Wahs, et al., Life Sciences, 31:637 (1982); See-m~n, et al., TIPS 15:264 (1994); Brewster, et al., J. Med. Chem., 33:1756 (1990); Sunahara, et al., Nature, 347:80 (1990); Chio, et al., Nature 343:266 (1990). In one embodiment, the Dl agonist is a selective agonist for the Dl .~e~lor over the D2 receptor (e.g., the compound has a lower Ki or EC50 for the D, receptor than the D2 receptor). In a further embo-lim~nt7 the Dl agonist is a weak agonist (e.g., K; or EC50 of greater than 1 ,uM or lmM) or is not a D2 agonist (e.g., Kj or EC50 of greater than 10 mM).
The Dl do~ h~e agonist may be any one or more of those substances known to those skilled in the art that are capable of activating or pu~P.~ g Dl do~al"h~e receptors. The Dl agonists that are suitable for use in the present invention include SKF38393, dihydrexidine, SKF 75670, SKF 82957, A77636, A68930, SKP
82526 (fenoldopam), and r~cemic trans-10, ll-dihydroxy 5, 6, 6a, 7, 8, 12b-hexahydro, and those Dl agonists disclosed in the references cited herein. The ~refe,l~d Dl dopamine agonist is SKF 38393.
What is meant by a "D2 dQp~min~ agonist" or a "D2 agonist" is a compound which is capable of activating or pol~ g D2 dopamine receptors (e.g., D2, D2 short and D2 long lecel)lo,~, D3, and D4 d~all.ine receptors). See, e.g., . . .

W O 97146239 PCTrUS97110838 Buntow, et al., Nature 3365:783-787 (1988), Del Toso, et al., EMBO J. 8:4025-4034 (1989), Giros, et al., Nature 342:923-926 (1989), Giaudy, et al., Proc. Nat. Acad.
Sci. U~ 86:9762-66 (1989), and Monsma, et al., Nature 342:926-929 (1989). In oneembo-limPnt~ the D2 agonist is a selective agonist for the D2 receptor over the D
5 leceylor. In a further embo~limPnt7 the D2 agonist is a weak Dl agonist or is not a D
agonist.
The D2 agonists for use in the present invention can be any one or more of those compounds known to those skilled in the art that are capable of activating or pole.,l;~l;n~ D2 dopamine receptors. D2 agonists suitable for use in the present10 invention include bçn"-",ides (e.g., sulpiride or raclopride), bulyl~henones (e.g., s~il.)pelidol) LY-171555, bromocriptine meth~nP sulfonate (+)-, 2,10,11-trihydroxyapomorphine HBr, R(-)-, lisuride hydrogen m~ te, 2-OH-NPA HCl, R(-)-, MDO-NPA HCl R(-), l~l~ylnol~anlorphine HCl R(-)-(NPA), 4uin~)irole HCl and those D2 agonists recited in the references cited herein.
A ~lt;rell~d class of D2 agonists includes ergot aLkaloids such as 2-bromo-alpha-ergocriptine (bromocriptine), dihydroef~o(a~ e, 6-methyl 8-beta-carbobenzyl~yaminoethyl-10-alpha-ergoline, 8-acylaminoergoline, 6-methyl-8-alpha-(N-acyl)amino-9-ergoline, pergoLide, licuri-le, 6-methyl-8-alpha-(N-phenyl-acetyl)amino-9-ergoline, ergocornine, 9,10-dihydroergocornine, any D-2-halo-6-alkyl-20 8-~ub~liluled ergoline, and D-2-bromo-6-methyl-8-cyanomethylergoline. Of these bromocriptine is most preferred.
Effective amounts of ergot aLkaloid for hl-m~nc and vellebldles when "i"i~t~,r~d alone (not conjoined to a Dl agonist) are typically within the range of 5.0 uglkg/day to 0.2 mg/kglday. However, the amount of the Dl agonist, or the D225 agonist, al agonist, a2 agonist, sel~ololml inhibitor, or SHT,B agonist depends on the condition being treated, the route of . ~ministration chosen, and the specific activity of the compound used and ultim~tP~ly will be decided by the ~ttenfling physician or In general, effective amounts of D2 agonist for hllm~ns and vellebl~tes 30 are within the range of 5 uglkglday to 3.5 mglkglday.
Three f~mili~s of ad~. er~ic receptors have been described in the CNS
(i.e., ~ 2, and ~B). See, e.g., Goodman & Gilman's The Pharmacological Basis of Therapeut~cs, ed. Hardman, et al. (9th ed. 1996). What is meant by an "adrenergic a WO 97146239 13 PCTrUS97/10838 antagonist" or an "a, antagonist" is a compound which is capable of blocking activation or down regulating cY~ ad~ er~ic rece~lols, e.g., binding to but not activating or down regulating ~see, e.g., Hieble, et al., J. Med. Chem., 38:3416 (1995);).
S Suitable assays for testing colnl~ullds for ~ or ~~l antagonist activity are well known in the art. See, e.g., Marshall, et al., Br. J. Ph~rmacol., 119:407 (1996);
C}~ n, J., J. Pharm. Ejcp. Ther., 264:375 (1993); Silva, et al., J. Pharm. Exp.
lher., 277:872 (1996); Paris, et al., Mol. Phann., 35:345 (1989); Morrow, et al., Mol. Pharm., 29:321 (1986); ~ndoin, et al., Life Sciences, 43:1805 (1988); Chein, et lO al., J. Med. Chem., 36:2196 (1993); Piercey, et al., Eur. J. Pharm., 317:29 (1996).

In one embodiment, the c~l antagonist is a selective antagonist for the c~, adl~ne~;ic receptor over the a!2 adrenergic receptor (e.g., the compound has as lower ~ for the cYl receptor than for the CY2 r~ce~tor). In another embo~lim~qnt the cr 15 antagonist is a weak a2 antagonist or is not a ~Y2 antagonist.
The ~x, antagonists for use in the present invention can be any one or more of those compounds known to those skilled in the art that directly or indirectly block activation of ~, adrenoceptors. The ~l antagonists suitable for use in the present invention include bromocriptine, benoxathin HCl, naftopidil 2HCl, (+)-niguldipine 20 HCl, S(~)-niguldipine HCl, prazosin HCl, doxazosin HCl, s~iperu-le HCl, urapidil HCl, 5-methyl urapidil, WB-4101 HCl, or those oll antagonists disclosed in the refelences cited herein.
Effective arnounts of a, antagonist for hum~n~ and ~iellel~ldles are generally within the range of 0.02 to 0.3 mg/kglday.
What is meant by an "a~llel e~ic ~2 agonist" or a "CY2 agonist" is a colll~uund capable of activating or ~ole~ g CY2 adrenergic receptors.
In one emboc~im~nt the ~~2 agonist is a selective agonist for the ~2 adrenergic receptor over the al adlenef~ic receptor. In another embodiment, the CY2 agonist is a weak ~, agonist or is not an ~l agonist.
The a~2 agonists for use in the present invention can be any one or more of those colnpou-lds known to those skilled in the art that are capable of activating C~2 adrenoceptors. The ~2 agonists suitable for use in the present invention includebromocriptine, epin~phrine~ no,~,~l~phrine, ~gm~tine sulfate, p-aminoc1Onidin~. HCl, W O 97/46239 14 PCTrUS97110838 B-HT 920 diHCl, B-HT 933 diHCl, clonidine HCl, guanabenz acetate, p-iodoclonilline HCl, oxymetazoline HCl, UK 14,304, and xylazine HCl or those C~2 agonists disclosed in the references cited herein.
Effective amounts of (X2 agonist for hllm~ns and v~ebldles are generally S within the range of 1 ug/kg/day to 0.3 mg/kg/day, and preferably between about 100 ug/kg/day and 0.25 mg/kg/day.
The se.uto~ ;ic inhibitors suitable for use in the present invention include bromocriptine.
Effective amounts of serotonergic inhibitors for hllm~n~ and ve,lebldles 10 are generally within the range of 5 ug/kg/day to 0.2 mg/kg/day.
When two (or more) agents are admini~tered in conjunction as disclosed in the Summary of Invention the amount of one or another can be lower than stated above, and even amounts that are subthreshold (when an agent is used singly) can be employed.
What is meant by a ''SHTIB agonist" is a compound which is capable of activating or pulç~l;AI;i~g SHT~B receptors. Suitable assays for testing compound for SHT,B activity are well known in the art. See, e.g., Schoeffer, et al., Naunyn-Schmiedeberg's Arch. Pharmacol., 339:675 (1989); Pauwels, et al., Eur. J.
Pharmacol., 290:95 (1995); Pauwels, et al., Neuropharm~cology, 33:67 (1994);
20 Parker, et al., J. Neurochemistry, 60:380 (1993).
In one embodiment, the SHTIB agonist is a selective agonist for the SHTIB receptor over the SHTIA receptor. In another embodiment, the SHTIB agonist is a selective for the SHT,B receptor over the 5HT2 receptor.
The SHT,B agonist may be any one or more of those substances known 25 to those skilled in the art that are capable of activating or potenti~ting SHT~B receptors.
Specific SHTIB agonists that are suitable for use in the present invention include RU24969 and CP93,129.
In general, in the practice of the invention, effective amounts of SHTIB
agonists for hllm~n~ and ~/ellebl~les are from about 0.01 to about 5.0 mg/kg of body 30 weight.
The dolJa~ e Dl agonist and the dopamine D2 agonist and/or other agent conjoined with the D, agonist (or with the D2 agonist) as well as the SHT,B
agonist may be ~lminictPred to a subject preferably orally, or ~ nleldlly, e.g., by subcut~neous, intravenous or intrAm~scular injection. Dermal delivery systems, e.g., skin patches, such as ionophoretic patches, as well as suppositories and other well-known systems for ~dminictç~ing ~h~ eutic~l agents can also be employed.
Sublingual, nasal and other tr~ncmucosal modes of ~minictration are also S conlt;---l)l~tçd ~ccPl~ ~,.IPd release compo.citions~ such as those disclosed in U.S.
Patent Application Ser. No. 08/459,021, are prerclled Each of the D2 agonist, ~l antagonist, ~2 agonist, serotonergic inhibitor, and SHT,B agonist are preferably ;~minictered at a predetermined time of day. The reason is that the effect of each of these agents on lipid and/or glucose metabolism is time-sensitive, as is explained in more detail for D2 agonists in U.S. Patent 5,585,347 and USSN 08/456,952, but is also applicable to the ~x, antagonists, a!2 agonists, serotonergic inhibitors, and SHTIB agonists. The l)r~fe,l~d time of ~1minilstration of Dl agonists, ~l antagonists, CY2 agonists, and serotonergic inhibitors is within an interval that results in effective blood levels of the agent(s) at a time during which the 15 standard prolactin levels in healthy subjects of the species to be treated are low. For example in hllm~nc standard prolactin levels are low between the hours of 7:00 and 22:00. Accordingly, the predrl~..lined time of ~dminictration of one or more of the foregoing agents is between the hours of 5:00 and 13:00, preferably between about 7:00 and about 12:00 or between 07:00 and 09:00. Divided doses can be ~(lminictered 20 and the schedule of :~dminictration can be varied to take into account pharmacokinetic pr~ellies of each active agent. Details of arlminictration are given in U.S. Patent 5,585,347 and USSN 08/456,952 for bromocriptine, but also apply to the ~~l antagonists, 1:~2 agonists, and seruloller~ic inhibitors employed in the present invention.
For mice the pr~felled time of ~lminist~tion of the first active agent is 25 within 1 hour after light onset. It is further pl~îerr~d that the ~lminictration take place when the subject is neither active nor feeding. The preferred time of . ~1minictr~tion for the second active agent, a SHT,B agonist, to mice is l l hours after light onset.
For other ~/elleb,al~ anim~lc the preferred time of administration of Dl, D2, ~ !2, and sGrulonel~ic inhibitors can be ascertained by reference to the standard 30 prolactin rhythm for the species of the animal to be treated. The standard prolactin curve can be gener~t~d by mç:~c--ring prolactin in young, healthy members of thespecies over a 24 hour period. The Dl, D2, al, ~X2, and stlolone~~ic inhibitors should CA 022~7084 1998-12-02 be :~minict~red at a time when prolactin levels are low in young, healthy, lean ~nim~1 of the same species and sex. See U.S. Patent 5,585,347 and USSN 08/456,952.
The, 1mini~tr~tion of the D~ agonist is also preferably timed, i.e. the D
agonist is also ~f1mini~tered at a prede(~ ecl time. Rec~lse the D, agonist amplifies the effect of the conjoined agent, it is advantageous to ~ l the Dl agonist at or about the time of ~ tion of the conjoined agent(s), such that the activity period of the D, agonist in the bloodstream of the treated subject overlaps (in fact preferably overlaps as much as possible) with the activity period of the conjoined agent. For conveMi~n~e of ~imini~tration and in order to promote subject compliance, the D
10 agonist can be ~iminiAtered at the same time as the conjoined agent(s).
The Dl agonist may but need not be in the same formulation or dosage form (or form part of the same composition) as the conjoined agent(s). If more than one conjoined agent is ~lminictered~ the conjoined agents may but need not be in the same formulation or dosage form or form part of the same composition.
In ll~a~ g human subjects, the SHTlB agonist is preferably a(iministered at night, most preferably at or shortly before a subject's bedtime. In other vertebrate ~nim~1~, the SHTIB agonist should be ~imini~tçred at or shortly before the time interval during which young, healthy, lean ~nim~1~ of the same species and sex have the highest daily levels of prolactin.
In treating vellebl~tes, generally, dosages of the Dl and D2 agonists and conjoined agent(s) are typically ~lmini~tered over a period ranging from about lO days to about 180 days, or longer~ Some p~ti~nt~ (e.g., patients in particularly poorphysical condition, or those of advanced age) may require a longer, or even continuous treatment, e.g., (i) until the condition (e.g., obesity, hy~e~ )idemia, Syndrome X, or 25 Type II diabetes) is nonn~li7e~ or (ii) for the lifetime of the patient. A tre~tm.q.nt duration e7~ce~1ing six months or even continuous treatment may be desirable even when not required. The SHTIB agonist ~timini~tr~tion will typically continue for as long as the Dl or D2 agonist ~mini~trAtion continues.
At least one of body fat deposits, body weight, plasma or blood glucose, 30 circu1~ting insulin, plasma triglycerides (TG), plasma free fatty acids (FFA), and food conb.~ll,plion of the subject will be reduced as the result of the treatment. Disorders of lipid and glucose metabolism are thereby treated and subjects s.lrreriilg from such pathologies as hyl~el~hagia, obesity, insulin resi~t~n-e (impaired glucose tolerance), W O 97/46239 17 PCT~US97/10838 hyperlipidemia, hyperinculin~mi~, and hyperglycemia will exhibit improvement in co.le~olldillg metabolic indices.
While a~l"vp~ ely timed ;3~lminictration of certain D, or D2 agonists (i.e., bromocriptine) alone will produce the effects described above to some degree, 5 these effects are ~mI)lified (pote~ d) by the conjoined ~lmini~trptir~n of the D, or D2 agonist agents and 5HTlB agonists ~tes~rihed in the present invention. In other words, the synergistic effect of the conjoined ~-lminict~tion of the Dl or D2 agonist and the conjoined SHT,8 agonist produces results that frequently are sllp~ric r to thoseexperienced through ~(lminictrati~n of the same amount of a D, or D2 agonist alone. It 10 should be noted that the present invention ptllllils but does not require each agent to be admini.~t~red in an amount over the threshold amount (in the absence of a conjoined agent) to improve one or more metabolic indices precisely because of the ~ngm~nted effect on these indices achieved by conjoined a~mini~t~tic)n according to the present invention.
These and other features of the invention will be better understood by reference to the ~li llents described in the examples below, which are inten-1ed to illustrate, but not in any way limit, the scope of the invention.

Difrer~l,l groups (8 mice per group) of 4-8 week old C57BL/6 ob/ob mice (lacking a functional leptin protein) were treated with:
1) vehicle at 1 and 11 hours after light onset (HALO);
2) Buspirone ("Busp"; a 5HTlA agonist) (3 mg/kg) at 11 HALO;

3) DOI (a SHT2 agonist) (8 mg/kg) at 11 HALO;

4) RU24969 ("RU") (SHTIB agonist) (3 mg/kg) at 11 HALO;

5) bromoc~i~Linc ("BC") (12mg/kg BW) at 1 HALO;

6) BC (12 mg/kg) at 1 HALO plus Buspirone (3 mgfkg) at 11 HALO;

7) BC (12 mg/kg) at 1 HALO plus DOI (8 mg/kg) at 11 HALO;

8) BC (12 mg/kg) at 1 HALO plus ~U (3 mg/kg) at 11 HALO;

9) SKF38393 ("SKF") (Dl agonist) (20mg/kg BW) at 1 HALO; and 10) SKF(20 mg/kg) at 1 HALO plus RIJ (3 mg/kg) at 11 HALO
for two weeks. Animals were held on 12-hour daily pholo~eliods and allowed to feed ad libitum. Food consumption was monitored daily for 3 days before the initiation of CA 022~7084 1998-12-02 W O 97/46239 18 PCT~US97/10838 treatment throughout the 14-day treqtm~nt period. Animals were sacrificed between 1 and 3 HALO on the day following the final treatment and plasma was collected for the analyses of insulin, glucose, and lipids while the carcasses were solubilized inethanolic KOH and analyzed for protein and lipid content. Blood glucose was S measured with an Accu-Chek Advantage glucose meter (Boeh. ;,-gel~). Serum insulin was measured with a r.q.-lini....-.ui-sqc.~qy kit (Linco Research) using rat insulin standards. Total triglycerides and free fatty acids were measured with kits from Sigma Diagnostics and Wako Ch~micql~, respectively. Body weight gain or loss as a result of treatment is depicted in Figure 1. Fat body mass is depicted in Figure 2. Lean 10 body mass is shown in Figure 3. Food consumption over the test period is shown in Figure 4. Blood glucose at the end of trç~tment is shown in Figure 5. Serum free fatty acids at the end of the treq-tment period is shown in Figure 6.
Bromocriptine, Buspirone, and DOI, individually, were ineffective in redl1cing body weight gain, body fat stores, or food intake. SKF and RU, 15 individually, caused a .cigni~ qnt decrease in body weight and food consumption.
Relative to controls, the body weight gain in BC/RU treated ~nimql~ was decreased by about 70%, fat body mass ~ignific~ntly decreased, lean body mass was not ~ignificantly decreased, and average daily food consumption decreased significantly, by about 28%.
Blood glucose was also signifi~ntly decreased relative to controls, as was serum free 20 fatty acids. (P < 0.05).
Treatment of mice with SKF and RU (SKF/RU) showed the most dramatic results. Food consumption was decreased by 52% and body weight showed a4g loss vs. a 6.4 g gain in controls. Body fat stores were decreased 32% compared to controls. Serum glucose was reduced by 42 %, and serum free fatty acids were 25 reduced 47 % .
Therefore, relative to control mice, the BC/RU and SKF/RU treated ~nim~lc con~l-med less food but did not decrease protein mass, while concu"cl,lly losing weight and fat. These data in-lic~te that the interactive effects of BC and RU or SKF and RU effectively reduced hyperphagia, obesity, hyperglycemia, and 30 lly~cl~idemia in the ob/ob mouse.

W O 97/46239 PCT~US97/10838 Dilr~e.ll groups of rats were injected ~ liloneally with SKF38393 ("SKF") (lOmg/kg BW) and bromocriptine ("BC") (lOmg/kg BW), SKF and RU24969 ("RU")(3 mg/kg BW), BC and SKF77434 (lOmg/kg BW), or vehicle for 8 days. The S BC and SKF were ndmini.~tered at 1 hour after light onset (HALO) and the RU and SKF77434 were atlminict~red at 11 HALO. Animals were held on 12-hour daily photoperiods and allowed to feed ad libitum. Food consumption was monitored daily for 3 days before the initi~tion of tre~tment throughout the 8-day treqtmçnt period.
The body weight of the rats at the beginning of the treatment period was from about 385 to 390 grams.
The combined tre~tment of bromocriptine and SKF38393 (BC/SKF) resulted in a decrease in food cons~ l"ption by 29% and a reduction in body weight of 15 grams (3.8%), whereas control rats gained an average of 27 grams over the same 8 day period (Fig. 7). An even more dramatic reduction in body weight was achievedby ~dmini.~tr~tion of BC and SKF77434 (a 6.2 ~ weight reduction) and SKF and RU
(an 8.2% weight reduction). Similarly, food consumption by BC/SKF treated rats decreased by 29 % over the 8 day period, but even greater reductions of 30%
(SKF/RU) and 46% (BC/SKF77434) were achieved by ;l~ministration of Dl or D2 agonist at 1 HALO and a SHTlB agonist at 11 HALO (Fig. 8). These data inr~ic~t~
20 that the interactive effects of Dl or D2 agonists SHTlB agonists given at predetel,llined times are highly effective in the reduction of obesity and food consumption.

Claims (21)

What is claimed is:

1. A method for modifying or regulating at least one of glucose or lipid metabolism disorders, body fat, or body weight which comprises (a) administering at a first predetermined time interval to a human or vertebrate animal subject in need of such modification or regulation a member selected from the group consisting of D1 dopamine agonists, D2 dopamine agonists, adrenergic .alpha.1 antagonists, adrenergic .alpha.2 agonists, and serotonin inhibitors and (b) administering at a second predetermined time interval a 5HT1B agonist.

2. The method of claim 1 wherein said administrations are effective to decrease at least one of food consumption, body weight, body fat, plasma insulin, plasma glucose, plasma lipid, and plasma lipoprotein.

3. The method of claim 1, wherein said method comprises administering a D1 dopamine agonist.

4. The method of claim 2, wherein said method comprises administering a D1 dopamine agonist.

5. The method of claim 3 wherein the D1 dopamine agonist is SKF38393.

6. The method of claim 1, wherein said method comprises administering a D2 dopamine agonist.

7. The method of claim 2, wherein said method comprises administering a D2 dopamine agonist.

8. The method of claim 6 wherein the D2 dopamine agonist is an ergot alkaloid selected from the group consisting of 2-bromo-alpha-ergocriptine, 6-methyl 8 beta-carbobenzyloxyaminoethyl-10-alpha-ergoline, 8-acylaminoergoline, pergolide, lisuride, 6-methyl-8-alpha-(N-acyl) amino-9-ergoline, 6-methyl-8-alpha-(N-phenyl-acetyl)amino-9-ergoline, ergocornine, 9,10-dihydroergocornine, and D-2-halo-6-alkyl-8-substituted ergolines, D-2-bromo-6-methyl-8-cyanomethylergoline.

9. The method of claim 8 wherein the ergot alkaloid is bromocriptine.

10. The method of claim 1, wherein said method comprises administering a D1 dopamine agonist and a D2 dopamine agonist.

11. The method of claim 2, wherein said method comprises administering a D1 dopamine agonist and a D2 dopamine agonist.

12. The method of claim 10 which comprises adiministering the D1 dopamine agonist at about the same time as the D2 dopamine agonist.

13. The method of claim 11 which comprises administering the D1 dopamine agonist at about the same time as the D2 dopamine agonist.

14. A method for treating insulin resistance, obesity, or type II diabetes which comprises (a) administering at a first predetermined time interval to a human or vertebrate animal subject in need of such treatment a member selected from the group consisting of D1 dopamine agonists, D2 dopamine agonists, adrenergic .alpha.1 antagonists, adrenergic .alpha.2 agonists, and serotonin inhibitors; and (b) administering at a second predetermined time interval a 5HT1B agonist.

15. The method of claim 14, wherein said method comprises treating insulin resistance or type II diabetes.

16. The method of claim 14, wherein said method comprises treating obesity.

17. The method of claim 14, wherein said method comprises administering a D1 dopamine agonist.

18. The method of claim 14, wherein said method comprises administering a D2 dopamine agonist.

19. The method of claim 14, wherein said method comprises administering a D1 dopamine agonist and a D2 dopamine agonist.

20. The method of claim 9, wherein said D2 dopamine agonist is bromocriptine and the 5HT1B agonist is RU24969.

21. The method of claim 18, wherein said D2 dopamine agonist is bromocriptine and the 5HTIB agonist is RU24969.