CA2106765A1 - Cck analogs containing ó-substituted amino acids - Google Patents
Cck analogs containing ó-substituted amino acidsInfo
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- CA2106765A1 CA2106765A1 CA002106765A CA2106765A CA2106765A1 CA 2106765 A1 CA2106765 A1 CA 2106765A1 CA 002106765 A CA002106765 A CA 002106765A CA 2106765 A CA2106765 A CA 2106765A CA 2106765 A1 CA2106765 A1 CA 2106765A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1024—Tetrapeptides with the first amino acid being heterocyclic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
- C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
- C07C229/24—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having more than one carboxyl group bound to the carbon skeleton, e.g. aspartic acid
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- C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
- C07C271/06—Esters of carbamic acids
- C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
- C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C271/22—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
- C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
- C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
- C07C323/57—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups
- C07C323/58—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton
- C07C323/59—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton with acylated amino groups bound to the carbon skeleton
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
- C07D209/16—Tryptamines
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/595—Gastrins; Cholecystokinins [CCK]
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06104—Dipeptides with the first amino acid being acidic
- C07K5/06113—Asp- or Asn-amino acid
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1002—Tetrapeptides with the first amino acid being neutral
- C07K5/1016—Tetrapeptides with the first amino acid being neutral and aromatic or cycloaliphatic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/02—Ortho- or ortho- and peri-condensed systems
- C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
- C07C2603/06—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members
- C07C2603/10—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings
- C07C2603/12—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings only one five-membered ring
- C07C2603/18—Fluorenes; Hydrogenated fluorenes
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Abstract
The invention covers novel peptides which contain at least one .alpha.,.alpha.-disubstituted amino acid. The compounds are useful as agents in the treatment of obesity and gastrointestinal disorders associated with gastrin. They are also useful in treating gastrin-dependent tumors, or as antipsychotics. Further, the compounds are antianxiety agents, antiulcer agents, antidepressant agents, and are agents useful for preventing the withdrawal response produced by chronic treatment or use followed by chronic treatment followed by withdrawal from nicotine, diazepam, alcohol, cocaine, caffeine, or opioids. Also covered are processes for making the compounds, novel intermediates useful in their preparation, compositions containing them, and methods of using the compounds.
Description
WO 92~19253 PCr/US~2~03015 6 ~ 6 ~
CCK AN~LOGS CONTAINI~G oc--SUBSTITUTED AMINO ACIDS
Bi~C~CGROl~ND OF THE INV~NTION :
~
Ag~nts acting at c:entral cholecystokinin ~CCK) ::
receptors may induce satiety (Schiclc, Yak~h ancl Go, ~;
R2qulatorY Pept des 14: 277-291 (19B6) .
The CCR peptides are w~ dely distri}:~uted in variou~ organs of th~ bc:~dy including the 1 0 gastrointe~tinal txact ~ endocrine gla~ds ; 2uld the :;
nerves of .the peripheral and centr 1 ~er~rou~ ~yoetems.
ariou-~ biologically acti~r~ f orm~ ha~re been ide~ti~ied includi~g a 33-amino ac:id hormorie a~d ~arious car3:)oxyl-terminus fra~ments of this peptide (e. g., the octapeptide CCK26~33 ar~d the tetrap~ptide (CCK30-33).
~G. J. Dockray, Br. Med. Bull., 38 (No. 3) :253~258, 19~2).
The ~ariou~ CC~C p~ptides are thou~hl: to be ~-involved in the c:orltrol of ~mooth muscle `~
contractility, exoc i~e and endocrine gland secretionr .`~
sensory nerve transmission, and numerou~ brain function~. ~dministrati~n of the nati~e peptide~
cause gall bladder contraction, amyla~e ~ecretion, excitation of cent~a~ neuron~r inhibition o~ feeding, antico~vulsi~e actions, and other behavioral effects.
('ICholecy3tokinin: Isolation, Struc~ure and Functions," G~ ~. J. Glass, Ed., Ra~en Pre~s, New York, 1980, pp 169-221i J. E. Morley, ife Sciences 27:355-368, 1980; "Cholecystokinin in the Ner~ous System," J. de Belleroche and G. J. Dockray, ~d., Ellis Horwoodr Chichester, ~ngland, 1984, pp 110-127.) The high co~centrations of CCK peptides in many brain areas also indicate major brain functions for `~
the~e peptides (G. J. ~ockray, Br. Med. Bull., 38 WO 92/lg253 PCI/U~92/0301~
CCK AN~LOGS CONTAINI~G oc--SUBSTITUTED AMINO ACIDS
Bi~C~CGROl~ND OF THE INV~NTION :
~
Ag~nts acting at c:entral cholecystokinin ~CCK) ::
receptors may induce satiety (Schiclc, Yak~h ancl Go, ~;
R2qulatorY Pept des 14: 277-291 (19B6) .
The CCR peptides are w~ dely distri}:~uted in variou~ organs of th~ bc:~dy including the 1 0 gastrointe~tinal txact ~ endocrine gla~ds ; 2uld the :;
nerves of .the peripheral and centr 1 ~er~rou~ ~yoetems.
ariou-~ biologically acti~r~ f orm~ ha~re been ide~ti~ied includi~g a 33-amino ac:id hormorie a~d ~arious car3:)oxyl-terminus fra~ments of this peptide (e. g., the octapeptide CCK26~33 ar~d the tetrap~ptide (CCK30-33).
~G. J. Dockray, Br. Med. Bull., 38 (No. 3) :253~258, 19~2).
The ~ariou~ CC~C p~ptides are thou~hl: to be ~-involved in the c:orltrol of ~mooth muscle `~
contractility, exoc i~e and endocrine gland secretionr .`~
sensory nerve transmission, and numerou~ brain function~. ~dministrati~n of the nati~e peptide~
cause gall bladder contraction, amyla~e ~ecretion, excitation of cent~a~ neuron~r inhibition o~ feeding, antico~vulsi~e actions, and other behavioral effects.
('ICholecy3tokinin: Isolation, Struc~ure and Functions," G~ ~. J. Glass, Ed., Ra~en Pre~s, New York, 1980, pp 169-221i J. E. Morley, ife Sciences 27:355-368, 1980; "Cholecystokinin in the Ner~ous System," J. de Belleroche and G. J. Dockray, ~d., Ellis Horwoodr Chichester, ~ngland, 1984, pp 110-127.) The high co~centrations of CCK peptides in many brain areas also indicate major brain functions for `~
the~e peptides (G. J. ~ockray, Br. Med. Bull., 38 WO 92/lg253 PCI/U~92/0301~
2 1 ~ 6 1 ~ .j --2 ~
(No. 3) :253--258, 1~82) . The most abundant form of brain CCK f~und is CCK26-33, although small quantities of CCK30-33 exis~ (Rehfeld and Gotterman, J
Neurochem. 32 :1339--1341 ~1979) ) O The role o~ central nervous sy~tem C:CK is not known with certainty, but it has beerl implicated in th~ control of feeding (Della--Fera and Baile, ~as~ 206: 471--473 (1979~
Currently available Appetite suppr~ssant drugs either act peripherallyr by incr~a~ing energy expenditur2 (such as thyroxir~), or in some other marmer ~ such as the biguanide~s ), ox act by ~x~rting a central e. f~3ct on appetite or ~atiety.
R~cerltly patient with bule~ia ~ere sh~n t~ have lower than normal CC~C le~rels in their plasma (Geracioti, et al, ~ Journal of Yedicin~
319: 683 ~1988) ) . An additional role for CCK in the ~
periphery is to re~late th~ relea~e of ir~sulirl. CCK :-has been showsl to irlcrease the le~rels of in~ulin when admini~tered to mammals (~ushakoff, et al, J. Clin.
~0 ~ 65 395 (~9~7)). ~::
C terminal ~ra~ments of CCK ha~e recently been reported to function as CCK receptor antagonist (Jen~en, et al, ~ 757:250 ~1983)i Spanark~l, JO Biol. Chem. 258:6746 (1983)). Japane~e patent application 70/10506 to Miyao, et al, discloses a tetrapeptide derivati~ of the carboxy terminal sequence o~ gastrin (~ Trp-L-~y~-L-~sp-~Phe ~2) which has antigastrin activity. .
Centrally acting appetite ~uppressants either `~:
pot~ntiate central cat~ch~lamine pathways and tend to be stimulants (for example, amphetamine), or influence serotonergic pathway~ (~or example, fenflur~mine). ~-Other.forms of drug therapy include bulking agents which act by filli~g the stomach~ thereby inducing a "feeling" of satiety.
W092/19253 PCT/USg2/0301~
(No. 3) :253--258, 1~82) . The most abundant form of brain CCK f~und is CCK26-33, although small quantities of CCK30-33 exis~ (Rehfeld and Gotterman, J
Neurochem. 32 :1339--1341 ~1979) ) O The role o~ central nervous sy~tem C:CK is not known with certainty, but it has beerl implicated in th~ control of feeding (Della--Fera and Baile, ~as~ 206: 471--473 (1979~
Currently available Appetite suppr~ssant drugs either act peripherallyr by incr~a~ing energy expenditur2 (such as thyroxir~), or in some other marmer ~ such as the biguanide~s ), ox act by ~x~rting a central e. f~3ct on appetite or ~atiety.
R~cerltly patient with bule~ia ~ere sh~n t~ have lower than normal CC~C le~rels in their plasma (Geracioti, et al, ~ Journal of Yedicin~
319: 683 ~1988) ) . An additional role for CCK in the ~
periphery is to re~late th~ relea~e of ir~sulirl. CCK :-has been showsl to irlcrease the le~rels of in~ulin when admini~tered to mammals (~ushakoff, et al, J. Clin.
~0 ~ 65 395 (~9~7)). ~::
C terminal ~ra~ments of CCK ha~e recently been reported to function as CCK receptor antagonist (Jen~en, et al, ~ 757:250 ~1983)i Spanark~l, JO Biol. Chem. 258:6746 (1983)). Japane~e patent application 70/10506 to Miyao, et al, discloses a tetrapeptide derivati~ of the carboxy terminal sequence o~ gastrin (~ Trp-L-~y~-L-~sp-~Phe ~2) which has antigastrin activity. .
Centrally acting appetite ~uppressants either `~:
pot~ntiate central cat~ch~lamine pathways and tend to be stimulants (for example, amphetamine), or influence serotonergic pathway~ (~or example, fenflur~mine). ~-Other.forms of drug therapy include bulking agents which act by filli~g the stomach~ thereby inducing a "feeling" of satiety.
W092/19253 PCT/USg2/0301~
-3- 210 ~ I ~ 3 Wo 90/06937 ~overs derivatives of tetrapeptides a3 CCK agonists. The compounds are for treating gastrointestinll disorder~, central nervous system disorders~ i~sulin related di-Qorders, treatment of paln or regulating appetite.
SUMMARY OF T~E INVENTION
The in~ention relate~ to novel compounds w~ich are CC~ ligand~ and are ~,a-~ubstituted mono-, di-, tri-, tetra-, or pol~peptides of ~ormula : R7 R~ N ~ C' ~ N ~ C T
H O R4 R5 H Q R~
a~d ~c phar~ac~utically aeceptable salt~ th~reo~
wherein ~l, ~2, R3, R4, ~S, R6, R7, RB, R9, R-7, A~ and n are a~ d~fi~d below.
In commnn~y as~igned copending application : 07/690,755 filed April 24, l99l, a~d its continuation-in-part application 07~852,086 filed March 20, 1992, by Horwell, et al, the di~clssure of which is incorporat~d by reference, ~-sub~ituted polypeptides are disclo ed.
:The inYention al~ relates to a pharmaceutical compositio~ c~ntai~i~g an effecti~e amount of a compound according to formula I in combinatio~ with a pharmaceu~ieally acceptable carrier in unit do~ag~
form effecti~e for appetite suppres~ion.
Thë i~v~ntion further relates to a method o~
appetite suppre~sion i~ mammals which compri~es , adm~nisteri~g an amo~nt effecti~e to suppress appetite of the composition deQcribed abo~e to a mammal i~ need of ~uch treatment.
WO g~/t92S3 PCI~/US~/03015 2 1 0 6 7 6 ~ ~4 The in~ention further relates to methods of treating gastrointestinal disorder~, central nervous system disorders ~uch as CNS suppressants which can exhibit such effect as antipsychotic, neuroleptic, anxiolytic, and anticon~ulsant.
Th~ invention further relates to method~ of reducing gastric acid secretion arld. to treating gas~rointestinal ulcers. -~
The in~ention further relates to blocking the reacti~n cau3ed by withdrawal ~rom dxug or alcohol u e and to pot~ntiating the ~ff~cts of morphi~ a~d other ~:
opioids in treating pain. ~:
The in~ention further pro~id~ proces~e~ for the ~:~
preparation of compounds of fox~ula I.
The in~ention further pro~i~s novel intexmediates u~eful in the preparation ~f compounds of formula I and al~ pro~ides proce~s~s for the preparation of the intermediat~
BRIEF ~ESCRIPTION 0~ THE DR~W~NGS
Figure 1 shows Compound 2 is a partial agoni~t.
Figure 2 ~hows Compound 4 is an agoni~t and Compound 5 ;~
is a weak agonist.
Figure 3 shows Compound 6 is an agonist.
Figures 1 through 4 show CCK8s as agonist reference standards. ::
... :
D~STAIhED DESCRIPTION
. .
The f ollowing table provid~s a dictionary of ~he terms u~ed in the description of the in~vention~ -.
WO92~19~53 P~T/U~92~03015 _5_ 21067~ :
TABLE I .
Abbreviated Desi~nation GLY Glycine ~`
~-AL~ ~-Alanine GABA 4-Aminobutanoic Acid DAVA ~-Aminovaleric Acid ~;
MET L-Methionine ~'-TYR L-Tyrosine ASE ~-Aspartic Acid PHæ L-Ph~nylalanine j;.
TRP L~Tryptophan ~:
BOC Tertbutoxycarbonyl l-ADOC l-Adamantyloxycarbonyl 2-AD05 2 Adamant~loxycarbonyl Z Benzyl~ycarbo~yl F~OC 9-Fluorenylmethoxycarbonyl ~`
~ib a Aminoi~obutyric Acid ~-~
PFP P~ntafluoroph~nyl ;~
~, The compound~ of the pre~ant in~rention are analos~s of CCK 33-33 and CC:K 2 6-33 whiGh contain at least one resid~e of an amino acid which is oc, a-disubstituted. These compounds differ from the :~;
mammalian genetically coc~d na~ural peptides in that tha a sub~tituents cannot all simultanec>usly b~
hydroger~
The compounds of the present in~ention are 3 0 repreae~ked by the f o~nula , .. . . . ., .. , . ., .. ... , .. ,,, ~, .. , .. , ... ~ ..... . .. ... ..... ... . . . .
W~ g2/19253 PCI/IJS92/03015 ~10ti7~ 6-R3 1CH ) :-Rl--A--N C 'j;< N~C' ~
R9 : ;:
or a phar~s~aceutically acceptable ~alt thereof wherein ~1 i~ hydrogen, BOC, l P.doc, 2--Adoc, æ, FMOC, R12R13Rl4Si (CH2)m~t::O- ~rh~r~in ~ , R~3, and R14 are each independ~ntly lo~er allcyl of :E~om 1 :
to 3 carbon atom~ d m i~ an integ~r of from 0 to 3, R15Co or R15NHCo- whereiIl R15 ~-i~ a strai~t or branched alkyl of from 1 to 6 carbon~ or 1- or 2-adamantyl;
i5 a borld, Gly, B-~ GABA, D~V:~, ~ib, N~ (C~2) p whereirl p ilS an ir~teger of from ~o 6, --:
2 0 NHC:O ~
~et-Gly, Tyr--~et Gly, ~:
ASR- Tyr ~OSO3H), Asp-Tyr~t-Gly, Asp Tyr ~OSQ3H) ~et-Gly; :
R2, R4, R6, and R8 are each ind~pend~ntly hydrogen ;
with the pro~iso that not all are ~
simultaneously hydrogen, lowex alkyl, CEI--CH2, C_C~ CH2--CH--C~2, --CH2C_CH, --::H2Ar, 3~ ::H20R, --CH2O~r, --(CH2) qCO2R~ ICH2) ~N (Rj 2 .-. w~erein q i~ an islteger of f rom 0 to 3, R is hydrc: gen or lower alkyl, Ar is a mono~ or polyc:yclic unsubstitut~d or substituted car~o-- or heteroaromatic or hydroaromatic moi~ty;
WO 92J1~253 PCr/US92iû3015 .
_7_ 21 ~76~3 R3 is 2~ or 3 indolyl, 1- or 2~naphthyl, 2 or 3-benzothienyl, 2- or 3-benzofuranyl, 2 or 3-indazoyl or phenyl, any of the~e can have from 0 to 3 substitu~nts each independently s~lected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, methoxyr trifluorom~thyl, nitro, hydroxy, NH2, and OCF3;
~5 i~; lower alkyl, ~C~2~ rS ~C:H2) SCH3 wherein r is an irlteS~er of f rom 1 to 6 arld s is ~
irlt~g~r oiE from 0 to 6; . ~ ~.
n i~ t~ger of :rom 1 ~o 3;
:1~7 i B --OH, --COOH~
tetrazole, triazole, - COORl 6, _cONRloR~
wherein R16 is lower alkyl, and R10 a~d Rll ara as descxibed below;
R9 is el~c:ted from R3 abc~ve;
O
R17 is ~ ~, CH20R18 or ~I wh~rein R10 a~d Rll are each independently hydrc>gen, lower alkyl, alkoxyc:arbor~yl, carboxyalkyl, c~r R10 ~:
arld R11 together form a rins3 of from 3 ko 10 atoms with the ~itr~gen to w~ich thay are attac:hed which ring contains atoms sel~cted from carbon, nitrogen, o~yg~nr a~d ~ulfur;
3 0 and or CH2CO2H or C~2CONRlOE~ or CONE?.10 11 R and R3 R4 ~d R5, R6 and R7 ~ d R9 9 and R10 may together form a rir~g of from 3 to 8 atoms which atom~ are selecteà . rom carbon, nitrogen, oxygen, and sulfur.
~:"
WO 92/1g253 PCI/US92/03015 21 0 ~ 8-- ;
Preferred compounds of the instant invention are tnose of :Eormula I wherein Rl is Fmoc, Z, Boc, or Me3Si (CH2) 20CO~; :
A is a bOrldr Gly, B AI~, ~ib, Asp Tyr-Met--Gly, Asp-Tyr ~OS03H~ -Met Gly;
R2, R4, :R6, and R8 are each independ~ntly alkyl of from 1 to 3 carbon atc~ms, carboxyalkyl of 1 ` -or 2 s:arbon atoms/ or C~120H;
R3 is 3 indazolyl, 3--benzothieny~
2-benzofuranyl, 3-benzs::Euranyl, or 2-bromo-3-}:senzofuranyl;
R5 i~ i~obut l, ~e~c-butylethyl, n propyl I or i~propyl;
R7 is CONH2 / CO~Me2, or OH; ~ -~
R9 is selec:ted fxom R3;
R10 and Rll are each isldependently alkyl of from 1 to 3 c:arbon atom~ or tog~ther R10 and R
fs~rm a ring of from 4 to 8 atoms including the nitrogen to which they are attached.
~ore pref~rred com~?ound~ o~ the i~tant invention are those of formula wherein Rl i~ hydrcgen or BOC;
A is a bond or ~=ALA; `~
R2, R4, R6, and R8 are each indeperlclerltly CK3 or ~--C~2 2~;
R3 is 3-indolyl, l naphthyl, or 2-naphthyl;
R5 is n-butyl, n-pentyl, (CH2) 2SCH3 or (CH2) 2SCH2C:H3;
R7 is OH, tetrazole, triazole, or C02H;
3~ R9 is phenyl, l-naphthyl, or 2-naphthyl;
~10 and Rll are each indeplsndently hydrogen, C~I3 or together R10 a~d Rll fvrm a ring of 5 or 6 atoms including the nitrogen to whi~:h they are attac:hed . `~ `
-9- 21067~
~specially preferred compounds of the instant ~;
invention are s~lected from the group consisting of:
~-Methyl-DL-tryptophyl-L-methionyl L-a-aspartyl-L-phenylalani~amide, a-Methyl-D-trypt~phyl-L~methiony~L--~ aspartyl-L-phenylalaninamide, a-Methyl-~-tryptophyl-L-methionyl-~-oc-aspartyl~
phenylalani~amide, ~-~[(l,1-dimethylethoxy)-caxbonyl3-a-methyl~L~ -:
10 tryptophyl]-L-m~thionyl~L~a-aspartyl~
phe~ylalani~amide, N-t(1,1-dimethylethoxy)-c rbonyl] -methyl-DL-tryptophyl-~dmethionyl ~-a-a~par~yl~
phenylalaninamide, L-tryptophyl-2-methyl-DL~methionyl-~-a~aspartyl-~-phe~ylalaninamide, N-[(l,1-di~ethylethoxy)-carbonyl]-a-m~t~yl-D-tryptophyl-L-methionyl-L-a-aspartyl ~-phenylalaninamid~, Glycyl-a-methyl-D tryptophyl-~ methi~nyl-L-a-aspartyl-L~phenylalanin ~ide, Glycyl-a~methyl-D~-tryptophyl-L-methiony~
aspar~yl-~-ph~nylalaninamide, Glycyl-a~methyl-~-tryptophyl-~-m~thionyl-L-a~
aspartyl-~-phe~ylalaninamide, Trp-Met-Asp DL-MePhe-NH2 ~E3C02H, and Trp~Met-D~-~eAxp Phe-NH2~CF3C02H.
The D and the ~ configuration of the compounds of ~
formula I are possible at the chiral center~ and are ~:
included in the scope of the invention.
Pr~ferred compounds are tho~e wherein the -amino :~
acid residue are of the LL3 configuration.
The compounds of the pre3ent in~ention can have ~ultiple chiral centers d~pending on their ~tructures.
Center~ of asymmetry may exist on carbon atom~ bearing ,~
WO9~ 253 PCT/US9~/030~S
'~1067~
substituents Rl in formula I and at ~2, R3, R4, R5, R6, R7, R8, R9, and R17 in formula I. In particular, the compounds of the present invention m y exist as diast~reomers, mixtura~ of diastereomer , or as the mixed or the individual optical enantiomers. The present in~entlon contemplates all such ~orms of the compounds. The mixtures of diastereomers are typically obtained as ~ result of the reaction~
described more ~ully below. Indi~idual dia tereomers may ~ ~epar~ted from mixtures of the di.ast~r~omers by con~entional techniques ~uch a3 colu~n chro~atography -or repetiti~ r~cry~tallizatio~
The co~po~nd of the in~tant in~ention include the olvates and hydrates and phanmaceutically acceptable sa}ts of *ormula I.
The term lower ~lkyl means skraight or br~nched chain alkyl groups of f rom 1 to 6 carbon atoms unless oth~rwi~ specified. -.
The term alkoxycarbonyl means a carbon gro~p of from 1 to 4 carbon atoms.
~re~erred pha ~aceutically acceptable ~alts are benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, alumi~ ~, calcium, lithium, magn~sium, p~tassium, sodium, zinc, ~5 diethylamine, and tromethane.
Especially preferred phar~aceutically acceptable saltis are N-methylglucamine and xodium for acids and -HCl, sul~ate, and tri~luoroacetate for ba~s.
IndiYidual enantiomers may be ~eparated by convent~onal methods well known in the art such as con~eraion to a s~lt with an optically acti~e compound, ~ollowed by ~eparation by chromatography or recrystalli~ation and recon~ersion to the nonsalt form.
., .
W~ 92/192~3 PCr/US92/0301~
2 1 0 6 ~ ~S ~
The preferred stere~chemistry of the compounds of the inver~tion is that exhibited by the compour~d- of :~xample 2 . :~
The compounds of the present in~rention can be ~:
formed by coupling irldi~ridual substituted -amino acid~ by m~thod~ well known in the art. (See, for example, standard synthetic method~ discu~ed in the multi--~rolume treatise "The Peptides, Analysis, Synthesis, Biology, " by Gro~s a~ad ~einho~r, Aeademic Pre~s, New York. ) The indivic~ual s~absl:ituted alpha - amino acid starti~g mat~rials are g~ne~ally kx~oNn or, if not known, m~y b~ sy~th~siæed and, if desired, re~ol~red by methocls within th~ ~kill of the art.
(Synthesis of racemic ~DL3 a-methyl tryptophan methyl ester -- ~ee Bra~Sa, ~. F ., et al, ~DS:
17:829 (1980) ) .
The invention al~o include~ no~el interm~diate~ ~
which are u~e ul in the preparatio~ o~ the ~in~l proclucts. Th~se int~r~ediat~ are the~ following -:
compounds or the compourld on the olid pha~e resin;
for example, Fmoc--MePhe Resin.
Fmoc--MePhe ~H2 Fmoc-A,sp tOBut ) ~ ~Phe~
Fmoc--~qek ~p (OBut)--lePhe NH2, Fmoc:--Trp--~et-A~p (OBut)--MePhe--NH2, Asp ( OBut )--MePhe--NH2 ~
AXp--~gePhe--NH2, ,:~, Fmoc-Met-~sp-MePhe NH2, ~et~Asp(OBut)-MePhe-NH2, Met-Asp-MePhe-NH2, ~ Met-Asp(OBut)-MePhe-N~2, Fmoc-Trp-Met-A p-~ePhe-N~2, Fmoc-MeAsp(QBut)-Phe-N~2, Fmoc-MeAsp Phe-N~2, . ~.
Fmoc-~et-~eAsp(OBut)-Phe ~H2, W~92/19253 PCTJU~g2~0~15 21067~ 12-Fmoc-Trp ~et-MeAsp(OBut)-Phe-NH2, ;~
MQAsp~OBu~)-Phe-NH2, :~.
MeAsp--Phe~NH2, ~-Met MeAsp(OBut)--Phe--NH2r Met- MeAsp-Phe-NH2 r Fmoc--Met--MeA~p-Phe--NH2, Fmoc--Trp-Met--~qeAsp--Phe--NH2, MeMet Asp(OBut)-Phe-N~2, Fmo~-~eMet-Asp(OBu~-Phe-NH2, Fmoc-~eMet ~3p-Phe-~2 - ~ MeMet-~p-Phe-NH2, : :Fmoc-Trp-MeMet ~p(OBut)-~he-N~2, Fmoc Trp-~t-Asp Ph~-N~, Fmoc-~eTrp-~et-A~p(OBut)-Phe-NH2, Fmoc-MeTrp-Met-Asp-Phe-~H2, ~-Fmoc-Gly-T~p-~et-~sp MePhe-NH2, Fmoc-Gly-Trp-~et~p~OBut)MePh~ N~2 Gly-~rp-~t-~p (o~sut ) MePhe N~2, ~moeo&ly-Trp-~et-P~p-Phe~2~2, Fmc~c Gly-~ ~et~Ma~p (013ut) -Phe ~H2, Gly~Trp-Me~-Me~sp(OBut)-Phe NX2, Fmoc-Gly-Trp-M~et-A~p-~he-NH2~ :
Fmoc-Gly-Trp~e~ p(OBut) Phe~N~2, Gly~Trp-MeM~t-~sp(OBut) Phe-N~2, Fmoc-Gly ~eTrp-Met-A-~p~Phe-N~2, Fmoc-Gly-~eTrp-Met-Asp~OBu~)~Phe N~2, Gly-MeTrp-Met-Asp(OBut)-Phe-N~2, Fmoc-MeMet-OH, . Fmoc-MeMet OPFP, MeAsp(OBut)OH, F~oc-MeAsp~OBut)O~, and Fmoc-~eAsp(OB*)O~FP. ~:
For preparing pharmaceutical compositions from the compounds of this inve~tion, inert, ~-pharmaceutically acceptable carriers can be either `:
WO 9~/lg253 Pcr/US92/û30~5 -13~ 76~
solid or liquid. Solid form preparations inc:lude powders, tablets, dispersible granules, capsules, cachets, and suppositories.
~ solid carrier can be one or more substarlce~
which may al~o act a dilue~ts, fla~oxing agents~
solubilizers, lubricants, suspending agents, binders, or tabl~3t disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in a mixture with the f inely di~ided a . ti~e component . In tablets, the acti~Te component is mixed with th~ carrier ha~ring the n~ce~gary binding proper~ies in ~uitable proportic~ns and com~ac~ed in th~ shape a~d size de~ir~d.
For preparing ~uppository preparations, a low-melting wax such as a mixt:ure of fatty acid glyceride~ a~d cocoa but'cex is f irst melted and the active ingr~3dient is disper~ed therein by, ~or exampler s,'tirring. The molteIl homogeneous raixture is then poured into converlient s:ized molds and allo~ed to cool and solidify.
The powders and tablets preferably ccntain 5 to about 7 9% of t~e active cc~mponent . Suitable carriers are magn~sium carbonate, magnesium stearate, talc, lactose, sugar, pectin, de~trin, starch, tragacanth, methyl cellulosel ~odilLm carboxymethyl celluloQe, a low-melting wax, cocoa butter, and the like. :
Tha t~rm "preparation" i~ intended to include the . .
f ormulation of the acti~re component with enc:apsulating material as a caxrier providiny a capsul~ in which the 33 active c~mp~nent (with or without other carrier~:) is surrou~ded by a carxier which is thus in as~ociation with it . Similarly, cachets are ~ ncluded.
Tab- ets, powders, cachets, a~d c:apsules can be ~ .
u~ed as solid do~ag~ fc)rsns suitable for oral administration.
W~92/19253 P~T/US92/03015 2 1 ~ 6 7 ~
~iquid form preparations include ~olutions, ~uspensions, and emulsions. Sterile water or water-propylene glycol solutions of the acti~e compounds may be mentioned as a~ exa~ple of liquid preparations suitable for parenteral administration.
Liquid preparations can also be ~ormulated in solution in aqueous polyethylene glycol ~olution. ~:-Aqueous s~lutions for oral administration can be prepared by di~solving the acti~e component in water and adding suitable color~nts, flavoring agen~s, ~tabilizers, a~d thickening agents as desir~d.
~queous su~p~nsio~s for oral u~e can be made by dispersing the finely di~id~d a~ti~e componen~ in water together with a ~iQCous material such as natural synthetic ~ums, resins, methyl cellulose, sodium carboxymethyl cel}ulo~e, a~d other susp~nding ~gents known to the pharmaceutical fo~mulation art.
Preferably, t~e pharmaceutical preparation is in ~:
unit d~sage f orm. In Quch foxm, the preparation i~
di~ided into unit doses containing appropriat~
quantities of the acti~e r~mponent. The unit dosage form can be a packaged preparation, the packag~
co~tai~ing discrete quantitie~ of the preparation, for example, packet~d tablets, cap~ules, and po~ders in ~ials or ampoules. The unit do~age form c~n al~o be a ` :~
capsule, cachet, or tablet it~elf, or it can be the ~-appropriat~ number of any of the~e packaged f~rms.
The biological activity of compounds of the ~.
present inventio~ was e~aluated employing an initial .
screaning test which rapidly and accurately measured the bi~ding of the tested co~pound to be known CCK
receptor sites. Speci~ic CCK r~ceptors have been shown to exist in the central nervous system. (See Hays et al, NeuropePtides 1:53-62 (1980); and Satuer ~t al, Science 208:11555-56 (1980).
WO ~2/192~3 ~/US92/0301S
--15 ~
In this screening test, the cerebral cortices taken from male CFL~? mice weighing between 30 to 40 g were di ssected on ice, weighed, and homogenized in 10 ~rolumes of 50 rn~ Tris--~ICl buffer ~pH 7 . 4 at 0-4 C) .
The resulting su~pension was centrifuged, the supernate was d:Lscarded, and the pellet ~as waQhed by resuspension in Tris--HCl i:uffer followed by recentri:Eugation. The final pellet was re~uspended in 20 volumes of lO nM Hepes buffer (pH 7 . 2 at 23t::) containing 130 r~ MaCl~ 4 . 7 nP~ KCl, 5 nM MgCl2, 1 r~q EDT~ 5 mg~mI bo~ine albumiI~, ar~d bacitracin . ~ -( O . 25 mg/snL) . - .;
In saturation studi~s, cex@bra~ cortical membranes were incubatad at 23C for 120 minutes in a :
final volume of 500 ,uliter of Hepes incubation buffer (pH 7 . 2) togather with 0 . 2-20 nM tritiated- -pentagastrin (Amersham International, Engl~nd~ .
In the di~3plac~ment ~xpe:riments, membra~es were `
incubated with a ~ gle concentration (2 ~ of : :~
ligand, toget~er with increa~ing conc~r~tration~ (lO ll to 1Q-l4M) of competitive test c:omp~und. In each case, the nonspecific bi~di~g was defined as that persisting ~`
in the pre ence of the unlabeled octapeptide CCK26_33 (10-6~) . , Following incubation, radioactivity boun~ to membranes was s~parated from that ~ree in solution by rapid filtration through Whatman GF/B filters and washed three time~ with 4 mL of ice cold Tris-HCl buffer. Filt~rs from samples i~cubated with tritiated-pentaga~trin were placed in polyethyle~e vials with 4 mL of qcintillation cocktail, and the radioactivity was e timated by liquid scintillation spectrometry (efficiency 47-52%).
The specific bi~ding to CCK r~ceptor sites was . 35 def ined as the total bound tritiated-pentagastrin WO 92/192~3 PCr/US92/0301~
21~676~
--~6~
minus the amount of tritiated-pentagastrin bound in the prese~ce of 10-6 octapeptide, CCR26_33.
Saturation curves ~or specific:
tritiated-pentagastrin binding to mouse cortical membranes were analyzed by the methods of Scatchard (Ann. New York Acad Sci . 51 : 660--72 (1949), and Hill (J. Physiol . 40 : ~V--VIII (1910), to pro~ide estimates for the maximum numb~r of binding sites ~B~aX) and the ~quilibrium dissociation constant (Ka)~
In dis~?la ::ement e~periments, inhibition Gur~es were analyzed by ei~her logit-lot plot~ or the iterati~re cur~e fitting computer program AI~FIT :
(D~Lean, Mun~on and };~dbard, 197 8 ) ~o pro~ide estimat~3s of the IC50 and nH ~apparent Hill 1~ coeffi::ient~ value ) . IC50 ~ralu~s were defined as the concentration of test compound required to produce 50%
inhibition of spec:i~ic: binding. 3 Th~ i~hibition constant (Xi) of the te~t compound wa~ then calculated according to the 2 0 t::heng-Pru of f equation: ;
~`'''"
IC50 ` ::
Ki = ~7~
:
wherein [L] is the concentration of radiolab~l and X
is the e~uilibxi~ dissociation constant.
The Ki/M ~ralues fc~r se~eral representati~re compounds of the pre~ent in~ention are present in Table I I ~
. ........................................................... ;
WO 92/lg253 P~/11!i92J03~15 210~76~
TABI,E II. Binding to Central CCK Receptors Comp;:und Ki~nM (n) (l) 7 . 16 ~ 1 . 95 g (~) 5.~4 (~.23-7.1~ 3 (3) (4) 7 . 80 (5 .56 ~ 12 . ~) 3 -(5) 9.20 (6. 86-11 . 8) ~ .
(~ 30.40 (2~.6-3S.5) 3 (7~ -:
- (n) -- Number of a~ays ~lectrophysiological Data - :~
Explanations and protocols for similar experiments to these can be found in J. ~ughes, Proc.
~e~ 87: 6728-6732 (l990i, and reference~
cited therein.
Fi~xe 1 3how the dose respon~;e curve *or CCK8s beiEor~ and ~urirlg expo~ux~ to coD~pousld (2~ and a cur~re for compound (2) alone. The curves repres~nt the total number of action p~tenl:ials c~cc:urring durlng a respon~e to CC:KRs or (2 ) a~d are plot-ed as counts (Y-a~ rer~us do~e (X--axis). The cur~res for compound (2) indi ::ate that l:his compound has partial .:
agorli~t type properti~s in this experiment, ~ is C::K-8s, ~=il~= is t2), and ... - .... is CCK-8s + (2) P (1 1~) Figurs 2 shows the do e respon~e cur~s f or 3}I-CCX and CCK4 analogs on the same neurone in the V~. The total rLunber of action potenkials occurring 31~ during.. a r~spon~3~ is plott~d as counts ~Y-axis) ver~us dose of ligas~d (X~axis~. C:ur~res :Eor the ~ction ~X*A) / (X+B) were fitted to the points using RS1 ~oftware (A = maximl~m possible response, B w EC50 f compound calculat~d from cur~re). The curves indicate WO 92/19253 P~r/US92/03015 210~76~ -18-that both ~4) and (5) have agonist-like proper~ies in this experiment. ==o-= is (5), ....Q.... i~ BH-CCK, is (4), -- -- -- is (1078.292444*X) /
(5.395304e-03+X), _ is (1149. 663878X*) t (0 . 010686~X), ...... is ~llOO*X) / (~+0 .47g53), -~
is (llOO*X) / ~X+0 . 014358), ..... is (O.S) sm~othed CCR, and o i s CCX .
In Fis~ures 3 and 4 the dose response curves f or CCK and for (4), (5), and (6? both before and after exposure to the potent C:CK 8 antagonist PD 134308 .
The tvtal nu~ r of ac:tion pot~ntials occurring during a response i~ plotted as counts ~Y-axis) ~rer~us do~e (X-axis) . Cur~e~ for the function ~X*A~ / (X+B) were fitted to the point~ using RSl software. TheQe cur~res indicate that compounds ~4), (5), and (&) ha~e C:CK ~ .
agoniQt-like properties which are antagc~nized by CCK-B
antagc>niQt ~ ', In l?igure 3 ~ is CCK, ==-== is ( 8 ), is (6) ~ PD 134308, --~ is (X*1126. 907666) / .
(X+9.879552~, _ is (1123*X~/(X~0.140626), and =====--= is (112'?*X) / (X+0.031476) .
Irl Figure 4 ~ C:CK, ==111== is (5), ........ ~
is (4), ------D--- - is (5) + PD 134308, and ~ is ~ .
(4) ~ PD 134308. .:~
Tables III and IV summarize the four figures. :~
WO 92/19253 P~US9~/~3015 -19- 2~ D67~
TABLE I I I
Compound Profile Figure I~
:
(2) Partial 1 --agonist , .
(4) Agoni~t 214 . 0 (5) Weak 2479 ~ 5 agonist . .
SUMMARY OF T~E INVENTION
The in~ention relate~ to novel compounds w~ich are CC~ ligand~ and are ~,a-~ubstituted mono-, di-, tri-, tetra-, or pol~peptides of ~ormula : R7 R~ N ~ C' ~ N ~ C T
H O R4 R5 H Q R~
a~d ~c phar~ac~utically aeceptable salt~ th~reo~
wherein ~l, ~2, R3, R4, ~S, R6, R7, RB, R9, R-7, A~ and n are a~ d~fi~d below.
In commnn~y as~igned copending application : 07/690,755 filed April 24, l99l, a~d its continuation-in-part application 07~852,086 filed March 20, 1992, by Horwell, et al, the di~clssure of which is incorporat~d by reference, ~-sub~ituted polypeptides are disclo ed.
:The inYention al~ relates to a pharmaceutical compositio~ c~ntai~i~g an effecti~e amount of a compound according to formula I in combinatio~ with a pharmaceu~ieally acceptable carrier in unit do~ag~
form effecti~e for appetite suppres~ion.
Thë i~v~ntion further relates to a method o~
appetite suppre~sion i~ mammals which compri~es , adm~nisteri~g an amo~nt effecti~e to suppress appetite of the composition deQcribed abo~e to a mammal i~ need of ~uch treatment.
WO g~/t92S3 PCI~/US~/03015 2 1 0 6 7 6 ~ ~4 The in~ention further relates to methods of treating gastrointestinal disorder~, central nervous system disorders ~uch as CNS suppressants which can exhibit such effect as antipsychotic, neuroleptic, anxiolytic, and anticon~ulsant.
Th~ invention further relates to method~ of reducing gastric acid secretion arld. to treating gas~rointestinal ulcers. -~
The in~ention further relates to blocking the reacti~n cau3ed by withdrawal ~rom dxug or alcohol u e and to pot~ntiating the ~ff~cts of morphi~ a~d other ~:
opioids in treating pain. ~:
The in~ention further pro~id~ proces~e~ for the ~:~
preparation of compounds of fox~ula I.
The in~ention further pro~i~s novel intexmediates u~eful in the preparation ~f compounds of formula I and al~ pro~ides proce~s~s for the preparation of the intermediat~
BRIEF ~ESCRIPTION 0~ THE DR~W~NGS
Figure 1 shows Compound 2 is a partial agoni~t.
Figure 2 ~hows Compound 4 is an agoni~t and Compound 5 ;~
is a weak agonist.
Figure 3 shows Compound 6 is an agonist.
Figures 1 through 4 show CCK8s as agonist reference standards. ::
... :
D~STAIhED DESCRIPTION
. .
The f ollowing table provid~s a dictionary of ~he terms u~ed in the description of the in~vention~ -.
WO92~19~53 P~T/U~92~03015 _5_ 21067~ :
TABLE I .
Abbreviated Desi~nation GLY Glycine ~`
~-AL~ ~-Alanine GABA 4-Aminobutanoic Acid DAVA ~-Aminovaleric Acid ~;
MET L-Methionine ~'-TYR L-Tyrosine ASE ~-Aspartic Acid PHæ L-Ph~nylalanine j;.
TRP L~Tryptophan ~:
BOC Tertbutoxycarbonyl l-ADOC l-Adamantyloxycarbonyl 2-AD05 2 Adamant~loxycarbonyl Z Benzyl~ycarbo~yl F~OC 9-Fluorenylmethoxycarbonyl ~`
~ib a Aminoi~obutyric Acid ~-~
PFP P~ntafluoroph~nyl ;~
~, The compound~ of the pre~ant in~rention are analos~s of CCK 33-33 and CC:K 2 6-33 whiGh contain at least one resid~e of an amino acid which is oc, a-disubstituted. These compounds differ from the :~;
mammalian genetically coc~d na~ural peptides in that tha a sub~tituents cannot all simultanec>usly b~
hydroger~
The compounds of the present in~ention are 3 0 repreae~ked by the f o~nula , .. . . . ., .. , . ., .. ... , .. ,,, ~, .. , .. , ... ~ ..... . .. ... ..... ... . . . .
W~ g2/19253 PCI/IJS92/03015 ~10ti7~ 6-R3 1CH ) :-Rl--A--N C 'j;< N~C' ~
R9 : ;:
or a phar~s~aceutically acceptable ~alt thereof wherein ~1 i~ hydrogen, BOC, l P.doc, 2--Adoc, æ, FMOC, R12R13Rl4Si (CH2)m~t::O- ~rh~r~in ~ , R~3, and R14 are each independ~ntly lo~er allcyl of :E~om 1 :
to 3 carbon atom~ d m i~ an integ~r of from 0 to 3, R15Co or R15NHCo- whereiIl R15 ~-i~ a strai~t or branched alkyl of from 1 to 6 carbon~ or 1- or 2-adamantyl;
i5 a borld, Gly, B-~ GABA, D~V:~, ~ib, N~ (C~2) p whereirl p ilS an ir~teger of from ~o 6, --:
2 0 NHC:O ~
~et-Gly, Tyr--~et Gly, ~:
ASR- Tyr ~OSO3H), Asp-Tyr~t-Gly, Asp Tyr ~OSQ3H) ~et-Gly; :
R2, R4, R6, and R8 are each ind~pend~ntly hydrogen ;
with the pro~iso that not all are ~
simultaneously hydrogen, lowex alkyl, CEI--CH2, C_C~ CH2--CH--C~2, --CH2C_CH, --::H2Ar, 3~ ::H20R, --CH2O~r, --(CH2) qCO2R~ ICH2) ~N (Rj 2 .-. w~erein q i~ an islteger of f rom 0 to 3, R is hydrc: gen or lower alkyl, Ar is a mono~ or polyc:yclic unsubstitut~d or substituted car~o-- or heteroaromatic or hydroaromatic moi~ty;
WO 92J1~253 PCr/US92iû3015 .
_7_ 21 ~76~3 R3 is 2~ or 3 indolyl, 1- or 2~naphthyl, 2 or 3-benzothienyl, 2- or 3-benzofuranyl, 2 or 3-indazoyl or phenyl, any of the~e can have from 0 to 3 substitu~nts each independently s~lected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, methoxyr trifluorom~thyl, nitro, hydroxy, NH2, and OCF3;
~5 i~; lower alkyl, ~C~2~ rS ~C:H2) SCH3 wherein r is an irlteS~er of f rom 1 to 6 arld s is ~
irlt~g~r oiE from 0 to 6; . ~ ~.
n i~ t~ger of :rom 1 ~o 3;
:1~7 i B --OH, --COOH~
tetrazole, triazole, - COORl 6, _cONRloR~
wherein R16 is lower alkyl, and R10 a~d Rll ara as descxibed below;
R9 is el~c:ted from R3 abc~ve;
O
R17 is ~ ~, CH20R18 or ~I wh~rein R10 a~d Rll are each independently hydrc>gen, lower alkyl, alkoxyc:arbor~yl, carboxyalkyl, c~r R10 ~:
arld R11 together form a rins3 of from 3 ko 10 atoms with the ~itr~gen to w~ich thay are attac:hed which ring contains atoms sel~cted from carbon, nitrogen, o~yg~nr a~d ~ulfur;
3 0 and or CH2CO2H or C~2CONRlOE~ or CONE?.10 11 R and R3 R4 ~d R5, R6 and R7 ~ d R9 9 and R10 may together form a rir~g of from 3 to 8 atoms which atom~ are selecteà . rom carbon, nitrogen, oxygen, and sulfur.
~:"
WO 92/1g253 PCI/US92/03015 21 0 ~ 8-- ;
Preferred compounds of the instant invention are tnose of :Eormula I wherein Rl is Fmoc, Z, Boc, or Me3Si (CH2) 20CO~; :
A is a bOrldr Gly, B AI~, ~ib, Asp Tyr-Met--Gly, Asp-Tyr ~OS03H~ -Met Gly;
R2, R4, :R6, and R8 are each independ~ntly alkyl of from 1 to 3 carbon atc~ms, carboxyalkyl of 1 ` -or 2 s:arbon atoms/ or C~120H;
R3 is 3 indazolyl, 3--benzothieny~
2-benzofuranyl, 3-benzs::Euranyl, or 2-bromo-3-}:senzofuranyl;
R5 i~ i~obut l, ~e~c-butylethyl, n propyl I or i~propyl;
R7 is CONH2 / CO~Me2, or OH; ~ -~
R9 is selec:ted fxom R3;
R10 and Rll are each isldependently alkyl of from 1 to 3 c:arbon atom~ or tog~ther R10 and R
fs~rm a ring of from 4 to 8 atoms including the nitrogen to which they are attached.
~ore pref~rred com~?ound~ o~ the i~tant invention are those of formula wherein Rl i~ hydrcgen or BOC;
A is a bond or ~=ALA; `~
R2, R4, R6, and R8 are each indeperlclerltly CK3 or ~--C~2 2~;
R3 is 3-indolyl, l naphthyl, or 2-naphthyl;
R5 is n-butyl, n-pentyl, (CH2) 2SCH3 or (CH2) 2SCH2C:H3;
R7 is OH, tetrazole, triazole, or C02H;
3~ R9 is phenyl, l-naphthyl, or 2-naphthyl;
~10 and Rll are each indeplsndently hydrogen, C~I3 or together R10 a~d Rll fvrm a ring of 5 or 6 atoms including the nitrogen to whi~:h they are attac:hed . `~ `
-9- 21067~
~specially preferred compounds of the instant ~;
invention are s~lected from the group consisting of:
~-Methyl-DL-tryptophyl-L-methionyl L-a-aspartyl-L-phenylalani~amide, a-Methyl-D-trypt~phyl-L~methiony~L--~ aspartyl-L-phenylalaninamide, a-Methyl-~-tryptophyl-L-methionyl-~-oc-aspartyl~
phenylalani~amide, ~-~[(l,1-dimethylethoxy)-caxbonyl3-a-methyl~L~ -:
10 tryptophyl]-L-m~thionyl~L~a-aspartyl~
phe~ylalani~amide, N-t(1,1-dimethylethoxy)-c rbonyl] -methyl-DL-tryptophyl-~dmethionyl ~-a-a~par~yl~
phenylalaninamide, L-tryptophyl-2-methyl-DL~methionyl-~-a~aspartyl-~-phe~ylalaninamide, N-[(l,1-di~ethylethoxy)-carbonyl]-a-m~t~yl-D-tryptophyl-L-methionyl-L-a-aspartyl ~-phenylalaninamid~, Glycyl-a-methyl-D tryptophyl-~ methi~nyl-L-a-aspartyl-L~phenylalanin ~ide, Glycyl-a~methyl-D~-tryptophyl-L-methiony~
aspar~yl-~-ph~nylalaninamide, Glycyl-a~methyl-~-tryptophyl-~-m~thionyl-L-a~
aspartyl-~-phe~ylalaninamide, Trp-Met-Asp DL-MePhe-NH2 ~E3C02H, and Trp~Met-D~-~eAxp Phe-NH2~CF3C02H.
The D and the ~ configuration of the compounds of ~
formula I are possible at the chiral center~ and are ~:
included in the scope of the invention.
Pr~ferred compounds are tho~e wherein the -amino :~
acid residue are of the LL3 configuration.
The compounds of the pre3ent in~ention can have ~ultiple chiral centers d~pending on their ~tructures.
Center~ of asymmetry may exist on carbon atom~ bearing ,~
WO9~ 253 PCT/US9~/030~S
'~1067~
substituents Rl in formula I and at ~2, R3, R4, R5, R6, R7, R8, R9, and R17 in formula I. In particular, the compounds of the present invention m y exist as diast~reomers, mixtura~ of diastereomer , or as the mixed or the individual optical enantiomers. The present in~entlon contemplates all such ~orms of the compounds. The mixtures of diastereomers are typically obtained as ~ result of the reaction~
described more ~ully below. Indi~idual dia tereomers may ~ ~epar~ted from mixtures of the di.ast~r~omers by con~entional techniques ~uch a3 colu~n chro~atography -or repetiti~ r~cry~tallizatio~
The co~po~nd of the in~tant in~ention include the olvates and hydrates and phanmaceutically acceptable sa}ts of *ormula I.
The term lower ~lkyl means skraight or br~nched chain alkyl groups of f rom 1 to 6 carbon atoms unless oth~rwi~ specified. -.
The term alkoxycarbonyl means a carbon gro~p of from 1 to 4 carbon atoms.
~re~erred pha ~aceutically acceptable ~alts are benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, alumi~ ~, calcium, lithium, magn~sium, p~tassium, sodium, zinc, ~5 diethylamine, and tromethane.
Especially preferred phar~aceutically acceptable saltis are N-methylglucamine and xodium for acids and -HCl, sul~ate, and tri~luoroacetate for ba~s.
IndiYidual enantiomers may be ~eparated by convent~onal methods well known in the art such as con~eraion to a s~lt with an optically acti~e compound, ~ollowed by ~eparation by chromatography or recrystalli~ation and recon~ersion to the nonsalt form.
., .
W~ 92/192~3 PCr/US92/0301~
2 1 0 6 ~ ~S ~
The preferred stere~chemistry of the compounds of the inver~tion is that exhibited by the compour~d- of :~xample 2 . :~
The compounds of the present in~rention can be ~:
formed by coupling irldi~ridual substituted -amino acid~ by m~thod~ well known in the art. (See, for example, standard synthetic method~ discu~ed in the multi--~rolume treatise "The Peptides, Analysis, Synthesis, Biology, " by Gro~s a~ad ~einho~r, Aeademic Pre~s, New York. ) The indivic~ual s~absl:ituted alpha - amino acid starti~g mat~rials are g~ne~ally kx~oNn or, if not known, m~y b~ sy~th~siæed and, if desired, re~ol~red by methocls within th~ ~kill of the art.
(Synthesis of racemic ~DL3 a-methyl tryptophan methyl ester -- ~ee Bra~Sa, ~. F ., et al, ~DS:
17:829 (1980) ) .
The invention al~o include~ no~el interm~diate~ ~
which are u~e ul in the preparatio~ o~ the ~in~l proclucts. Th~se int~r~ediat~ are the~ following -:
compounds or the compourld on the olid pha~e resin;
for example, Fmoc--MePhe Resin.
Fmoc--MePhe ~H2 Fmoc-A,sp tOBut ) ~ ~Phe~
Fmoc--~qek ~p (OBut)--lePhe NH2, Fmoc:--Trp--~et-A~p (OBut)--MePhe--NH2, Asp ( OBut )--MePhe--NH2 ~
AXp--~gePhe--NH2, ,:~, Fmoc-Met-~sp-MePhe NH2, ~et~Asp(OBut)-MePhe-NH2, Met-Asp-MePhe-NH2, ~ Met-Asp(OBut)-MePhe-N~2, Fmoc-Trp-Met-A p-~ePhe-N~2, Fmoc-MeAsp(QBut)-Phe-N~2, Fmoc-MeAsp Phe-N~2, . ~.
Fmoc-~et-~eAsp(OBut)-Phe ~H2, W~92/19253 PCTJU~g2~0~15 21067~ 12-Fmoc-Trp ~et-MeAsp(OBut)-Phe-NH2, ;~
MQAsp~OBu~)-Phe-NH2, :~.
MeAsp--Phe~NH2, ~-Met MeAsp(OBut)--Phe--NH2r Met- MeAsp-Phe-NH2 r Fmoc--Met--MeA~p-Phe--NH2, Fmoc--Trp-Met--~qeAsp--Phe--NH2, MeMet Asp(OBut)-Phe-N~2, Fmo~-~eMet-Asp(OBu~-Phe-NH2, Fmoc-~eMet ~3p-Phe-~2 - ~ MeMet-~p-Phe-NH2, : :Fmoc-Trp-MeMet ~p(OBut)-~he-N~2, Fmoc Trp-~t-Asp Ph~-N~, Fmoc-~eTrp-~et-A~p(OBut)-Phe-NH2, Fmoc-MeTrp-Met-Asp-Phe-~H2, ~-Fmoc-Gly-T~p-~et-~sp MePhe-NH2, Fmoc-Gly-Trp-~et~p~OBut)MePh~ N~2 Gly-~rp-~t-~p (o~sut ) MePhe N~2, ~moeo&ly-Trp-~et-P~p-Phe~2~2, Fmc~c Gly-~ ~et~Ma~p (013ut) -Phe ~H2, Gly~Trp-Me~-Me~sp(OBut)-Phe NX2, Fmoc-Gly-Trp-M~et-A~p-~he-NH2~ :
Fmoc-Gly-Trp~e~ p(OBut) Phe~N~2, Gly~Trp-MeM~t-~sp(OBut) Phe-N~2, Fmoc-Gly ~eTrp-Met-A-~p~Phe-N~2, Fmoc-Gly-~eTrp-Met-Asp~OBu~)~Phe N~2, Gly-MeTrp-Met-Asp(OBut)-Phe-N~2, Fmoc-MeMet-OH, . Fmoc-MeMet OPFP, MeAsp(OBut)OH, F~oc-MeAsp~OBut)O~, and Fmoc-~eAsp(OB*)O~FP. ~:
For preparing pharmaceutical compositions from the compounds of this inve~tion, inert, ~-pharmaceutically acceptable carriers can be either `:
WO 9~/lg253 Pcr/US92/û30~5 -13~ 76~
solid or liquid. Solid form preparations inc:lude powders, tablets, dispersible granules, capsules, cachets, and suppositories.
~ solid carrier can be one or more substarlce~
which may al~o act a dilue~ts, fla~oxing agents~
solubilizers, lubricants, suspending agents, binders, or tabl~3t disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in a mixture with the f inely di~ided a . ti~e component . In tablets, the acti~Te component is mixed with th~ carrier ha~ring the n~ce~gary binding proper~ies in ~uitable proportic~ns and com~ac~ed in th~ shape a~d size de~ir~d.
For preparing ~uppository preparations, a low-melting wax such as a mixt:ure of fatty acid glyceride~ a~d cocoa but'cex is f irst melted and the active ingr~3dient is disper~ed therein by, ~or exampler s,'tirring. The molteIl homogeneous raixture is then poured into converlient s:ized molds and allo~ed to cool and solidify.
The powders and tablets preferably ccntain 5 to about 7 9% of t~e active cc~mponent . Suitable carriers are magn~sium carbonate, magnesium stearate, talc, lactose, sugar, pectin, de~trin, starch, tragacanth, methyl cellulosel ~odilLm carboxymethyl celluloQe, a low-melting wax, cocoa butter, and the like. :
Tha t~rm "preparation" i~ intended to include the . .
f ormulation of the acti~re component with enc:apsulating material as a caxrier providiny a capsul~ in which the 33 active c~mp~nent (with or without other carrier~:) is surrou~ded by a carxier which is thus in as~ociation with it . Similarly, cachets are ~ ncluded.
Tab- ets, powders, cachets, a~d c:apsules can be ~ .
u~ed as solid do~ag~ fc)rsns suitable for oral administration.
W~92/19253 P~T/US92/03015 2 1 ~ 6 7 ~
~iquid form preparations include ~olutions, ~uspensions, and emulsions. Sterile water or water-propylene glycol solutions of the acti~e compounds may be mentioned as a~ exa~ple of liquid preparations suitable for parenteral administration.
Liquid preparations can also be ~ormulated in solution in aqueous polyethylene glycol ~olution. ~:-Aqueous s~lutions for oral administration can be prepared by di~solving the acti~e component in water and adding suitable color~nts, flavoring agen~s, ~tabilizers, a~d thickening agents as desir~d.
~queous su~p~nsio~s for oral u~e can be made by dispersing the finely di~id~d a~ti~e componen~ in water together with a ~iQCous material such as natural synthetic ~ums, resins, methyl cellulose, sodium carboxymethyl cel}ulo~e, a~d other susp~nding ~gents known to the pharmaceutical fo~mulation art.
Preferably, t~e pharmaceutical preparation is in ~:
unit d~sage f orm. In Quch foxm, the preparation i~
di~ided into unit doses containing appropriat~
quantities of the acti~e r~mponent. The unit dosage form can be a packaged preparation, the packag~
co~tai~ing discrete quantitie~ of the preparation, for example, packet~d tablets, cap~ules, and po~ders in ~ials or ampoules. The unit do~age form c~n al~o be a ` :~
capsule, cachet, or tablet it~elf, or it can be the ~-appropriat~ number of any of the~e packaged f~rms.
The biological activity of compounds of the ~.
present inventio~ was e~aluated employing an initial .
screaning test which rapidly and accurately measured the bi~ding of the tested co~pound to be known CCK
receptor sites. Speci~ic CCK r~ceptors have been shown to exist in the central nervous system. (See Hays et al, NeuropePtides 1:53-62 (1980); and Satuer ~t al, Science 208:11555-56 (1980).
WO ~2/192~3 ~/US92/0301S
--15 ~
In this screening test, the cerebral cortices taken from male CFL~? mice weighing between 30 to 40 g were di ssected on ice, weighed, and homogenized in 10 ~rolumes of 50 rn~ Tris--~ICl buffer ~pH 7 . 4 at 0-4 C) .
The resulting su~pension was centrifuged, the supernate was d:Lscarded, and the pellet ~as waQhed by resuspension in Tris--HCl i:uffer followed by recentri:Eugation. The final pellet was re~uspended in 20 volumes of lO nM Hepes buffer (pH 7 . 2 at 23t::) containing 130 r~ MaCl~ 4 . 7 nP~ KCl, 5 nM MgCl2, 1 r~q EDT~ 5 mg~mI bo~ine albumiI~, ar~d bacitracin . ~ -( O . 25 mg/snL) . - .;
In saturation studi~s, cex@bra~ cortical membranes were incubatad at 23C for 120 minutes in a :
final volume of 500 ,uliter of Hepes incubation buffer (pH 7 . 2) togather with 0 . 2-20 nM tritiated- -pentagastrin (Amersham International, Engl~nd~ .
In the di~3plac~ment ~xpe:riments, membra~es were `
incubated with a ~ gle concentration (2 ~ of : :~
ligand, toget~er with increa~ing conc~r~tration~ (lO ll to 1Q-l4M) of competitive test c:omp~und. In each case, the nonspecific bi~di~g was defined as that persisting ~`
in the pre ence of the unlabeled octapeptide CCK26_33 (10-6~) . , Following incubation, radioactivity boun~ to membranes was s~parated from that ~ree in solution by rapid filtration through Whatman GF/B filters and washed three time~ with 4 mL of ice cold Tris-HCl buffer. Filt~rs from samples i~cubated with tritiated-pentaga~trin were placed in polyethyle~e vials with 4 mL of qcintillation cocktail, and the radioactivity was e timated by liquid scintillation spectrometry (efficiency 47-52%).
The specific bi~ding to CCK r~ceptor sites was . 35 def ined as the total bound tritiated-pentagastrin WO 92/192~3 PCr/US92/0301~
21~676~
--~6~
minus the amount of tritiated-pentagastrin bound in the prese~ce of 10-6 octapeptide, CCR26_33.
Saturation curves ~or specific:
tritiated-pentagastrin binding to mouse cortical membranes were analyzed by the methods of Scatchard (Ann. New York Acad Sci . 51 : 660--72 (1949), and Hill (J. Physiol . 40 : ~V--VIII (1910), to pro~ide estimates for the maximum numb~r of binding sites ~B~aX) and the ~quilibrium dissociation constant (Ka)~
In dis~?la ::ement e~periments, inhibition Gur~es were analyzed by ei~her logit-lot plot~ or the iterati~re cur~e fitting computer program AI~FIT :
(D~Lean, Mun~on and };~dbard, 197 8 ) ~o pro~ide estimat~3s of the IC50 and nH ~apparent Hill 1~ coeffi::ient~ value ) . IC50 ~ralu~s were defined as the concentration of test compound required to produce 50%
inhibition of spec:i~ic: binding. 3 Th~ i~hibition constant (Xi) of the te~t compound wa~ then calculated according to the 2 0 t::heng-Pru of f equation: ;
~`'''"
IC50 ` ::
Ki = ~7~
:
wherein [L] is the concentration of radiolab~l and X
is the e~uilibxi~ dissociation constant.
The Ki/M ~ralues fc~r se~eral representati~re compounds of the pre~ent in~ention are present in Table I I ~
. ........................................................... ;
WO 92/lg253 P~/11!i92J03~15 210~76~
TABI,E II. Binding to Central CCK Receptors Comp;:und Ki~nM (n) (l) 7 . 16 ~ 1 . 95 g (~) 5.~4 (~.23-7.1~ 3 (3) (4) 7 . 80 (5 .56 ~ 12 . ~) 3 -(5) 9.20 (6. 86-11 . 8) ~ .
(~ 30.40 (2~.6-3S.5) 3 (7~ -:
- (n) -- Number of a~ays ~lectrophysiological Data - :~
Explanations and protocols for similar experiments to these can be found in J. ~ughes, Proc.
~e~ 87: 6728-6732 (l990i, and reference~
cited therein.
Fi~xe 1 3how the dose respon~;e curve *or CCK8s beiEor~ and ~urirlg expo~ux~ to coD~pousld (2~ and a cur~re for compound (2) alone. The curves repres~nt the total number of action p~tenl:ials c~cc:urring durlng a respon~e to CC:KRs or (2 ) a~d are plot-ed as counts (Y-a~ rer~us do~e (X--axis). The cur~res for compound (2) indi ::ate that l:his compound has partial .:
agorli~t type properti~s in this experiment, ~ is C::K-8s, ~=il~= is t2), and ... - .... is CCK-8s + (2) P (1 1~) Figurs 2 shows the do e respon~e cur~s f or 3}I-CCX and CCK4 analogs on the same neurone in the V~. The total rLunber of action potenkials occurring 31~ during.. a r~spon~3~ is plott~d as counts ~Y-axis) ver~us dose of ligas~d (X~axis~. C:ur~res :Eor the ~ction ~X*A) / (X+B) were fitted to the points using RS1 ~oftware (A = maximl~m possible response, B w EC50 f compound calculat~d from cur~re). The curves indicate WO 92/19253 P~r/US92/03015 210~76~ -18-that both ~4) and (5) have agonist-like proper~ies in this experiment. ==o-= is (5), ....Q.... i~ BH-CCK, is (4), -- -- -- is (1078.292444*X) /
(5.395304e-03+X), _ is (1149. 663878X*) t (0 . 010686~X), ...... is ~llOO*X) / (~+0 .47g53), -~
is (llOO*X) / ~X+0 . 014358), ..... is (O.S) sm~othed CCR, and o i s CCX .
In Fis~ures 3 and 4 the dose response curves f or CCK and for (4), (5), and (6? both before and after exposure to the potent C:CK 8 antagonist PD 134308 .
The tvtal nu~ r of ac:tion pot~ntials occurring during a response i~ plotted as counts ~Y-axis) ~rer~us do~e (X-axis) . Cur~e~ for the function ~X*A~ / (X+B) were fitted to the point~ using RSl software. TheQe cur~res indicate that compounds ~4), (5), and (&) ha~e C:CK ~ .
agoniQt-like properties which are antagc~nized by CCK-B
antagc>niQt ~ ', In l?igure 3 ~ is CCK, ==-== is ( 8 ), is (6) ~ PD 134308, --~ is (X*1126. 907666) / .
(X+9.879552~, _ is (1123*X~/(X~0.140626), and =====--= is (112'?*X) / (X+0.031476) .
Irl Figure 4 ~ C:CK, ==111== is (5), ........ ~
is (4), ------D--- - is (5) + PD 134308, and ~ is ~ .
(4) ~ PD 134308. .:~
Tables III and IV summarize the four figures. :~
WO 92/19253 P~US9~/~3015 -19- 2~ D67~
TABLE I I I
Compound Profile Figure I~
:
(2) Partial 1 --agonist , .
(4) Agoni~t 214 . 0 (5) Weak 2479 ~ 5 agonist . .
(6) Agonist 3524 . U .
CCK8s Agonist 1~45 . 4 :l --~''""' , The compounds were further provén to be agonists following experaments in which they ~exe challeng~d with ~he previ~usily de~cri~ed ~otent and selecti~e CCK-B antagonist, reference compound PD 134308 named ~R-(R*,R*)-4~l2-~3-(lH-indol-3-yl)-2-methyl~1-oxo-2 ~[~tricyclo r 3 ~ 3 .1.13-7~ dec-2-yloxy~carbonyl]amino~
propyl]aminoJ-l-phenethyl]amino~-4-oxobutanoic acid.
TABLE IV
C:ompound Graph(PD 134308) /s~M
(4) + ref~rence compour~d 44 . 67 (5) + refçrenc:e compound 43 ~ 02 ~6) ~ reference compouslcl 3 14 . 4 CCK8~ ~ refererlce compound -- 5 . 4 Data shows ~hat compound (4) is equipotent with CC~8s while compound (6) possesses slightly lower el~ctrophysiological activity and compound (53 has relatively low agonist acti~ity. Compound ~2) îs a partial agonist.
WO~2/192S3 P~T/VS92/03015 ~13~ 76j -20~
Compounds of the present invention are useful as appetite suppre~a~ts as based on the tests described hexei~below.
I~ the Palatable Diet Feeding a~say, ~dult male Hooded Lister rats weighing between 200-400 g are hou~d indi~idually and trained to ~at a palatable diet. This diet consists of Nestl~s sweetened condensed milk, powdered rat food, and rat water which was when blended together set to a firm consi tency.
Each rat is pre~Qnted with 20 to 30 g of the pala~bl~
- diet for 30 minutes per day during the light pha~e of the lght-dark cycle o~r a trai~ing period o~ S day~
The inta~ ~f palatable diet is mea~ured by weighing -~
the food container before and after the 30~minute `~
access period (limits of ac~.uracy O.l g). Care is taken to collect and correct for a~y spillage of the diet. R~t~ ha~e free acces~ to pellet food and water ~;
exc~pt during the 30-minut~ t~t period. ~
After the traini~g perivd, do e~re~ponQ~ ~urve~ ::
are construct~d ~or CCX8 and several repre~ntati~e compounds of the present in~ention (n - 8-lO xats per do~e le~el). ~PE50 ~a~ue~ ('95% confidence limits) are obtained for the anorectic effeats of th~3e compounds.
In thera~eutic u~e as appetite Quppression :~
ag~nts, the compounds of the in tant invention are admi~istered to the patient at dosage le~els o from about 200 to about 2800 mg per day. . ~.
As ago~ists the compounds of the instant in~ention ha~e therapeutic uti~ity as appetite suppr~ants and~ as antagonists, as agents in ~.
treating ga~trointestinal disorders, central ner~ous ~ystem di~order~ such as CNS suppressants which can ;~
exhibit such effect~ as antipsychotics, n~uroleptics, anxiolytics, and antico~ulsants.
WO 92/lg2S3 PCrJUS92/~3015 210~7&~
As antagonists, the compounds o~ the instant invention are expected to have utility in reducing gastric acid secretion, in raducing anxiety, in : - ~
treating gastrointestinal ulcers , in treating ~:
psychotic behavior, in blockis~g the reaction caused by withdrawal from drug, alcohol, cocain~3, benzodiazepine, diazepam, or nicotine, and in pot~3ntiating the e:Efects of morphine for treating pain .
CCK A~alo~s c:ontaining o~ znethyl amiIlo acid~ were mad~ by three me~hods. ~ethocl A is a solutioxl phase procedure ~herein the peptides were a~bled uging standard 301Util:~n pha~e protocols yielding ~u~ ly protected interrnediates or standard salt coupling which yield a~partic acid side-chain d~protected intermediates. Examples 1 and 2 below illustrate Method A.
Th~3 peptides c:a~ also be prepared by ~ethod B, a solid-pha~ method wherein peptid~3~ were con. tructed on solid~pha^~ rg~inc d ~igned to produc~ C-termirlal amides either by tre~tment of the resin with a~unonia in methanol s~r by direct clea~age of an appropriate~ y ~:ubstituted resin usin~ trifluoroacetic acid, with the reguired ~c~engerr, gi~ing the amides directly. The latter protoGol was u ad ~ith DuPont ~apidAmide~ or Nova Biochem No~a~yn KR~ resins either in a simple bubbler apparatus ~DuPo~t re~in) or automated synthesizer (Nova Biochem re~in). This method is illustrated by Example 3 b~low.
The third method, ~ethod C, is a non~ol~t method whereby the amino compo~e~t ~nd a pentafluoroph~yl e~ter ar~ heated together without a solvent at 50~ to 100C for lO to 120 minutes or more.
ThiQ method is illu~trated by E~ampl~ 4 b~low.
~ .
WO 92/192S3 PCr/US92J03015 21 1~ ~7 ~3 Male Hooded ~ister rats (175--2~0 g) are housed individually and fasted overnight (free accese to ` ~:
watex~ . They are arlesthetized with urethane ~1. 5 g/ky IP) and the trachea ca~ulated to aid ~pontaneous `:
respiration. The stomach i~ perfused conl;inuously using a modification of the original method of Ghosh & ~ ~
Schild in "Continuous recording of acid secretion in . ~' the rat", Br._J~aE~. 13:54--61, 1956 as described :~:
by Parsons in "Quantitative studi~3s of drug-inducecl gastric acid ~ecretion". ~Ph.D. Thesis, Uni~ersity of London, 1969? . Th~ ca~ity of the: ~tomach is per~u ed at a rate of 3 ~/min with 5.4% w~v gllleose ~olution ~:
throu~h both the e~phaç~eal and body cannula. ~he fluid is propelled by a roll~3r pump (Gilson, ~inipuls 2), through heating coils to bring its temperature to 37 ~t 1C. The perfusion ~luid i~
collected by the fundic collecting f~nnel and pas~ed to a p~ electrode connected to a Jenway pH met2r ~PE~6~. ~n output is taken from the p~ meter to a Rikadenki chart recorder f or the on~line recording of the p~ of the gastric perfu~ate.
Pentagastrin is stored as a frozen aliquot and diluted to the r~quired concentrations ~ith st~rile O . 9% w/~r NaCl . No~rel com~c~unds are diæ~3~1~ed in :
steril~ 0. 9% w/~r NaCl on the day of the experiment. ~:
Drugs are administered I'V through a canslulated jugular :
~rein s a bolus in a dose ~olume of 1 mL jkg wa~h~3d in with 0 .15 m~ O . 9% w/~ NaCl. Ba~al pH is allo~ed to ~`.
stabilize ~ef or~ administration o~ compourld~ is begun .
Typically 30 minutes elap~3es between surgery and the :Eirst .~ompound a~ninistration . ~:
Compounds of the in~rention may al~o ar~tagonize the stimulation of 5~astric ac:id secretion produt::ed by ~;~
a standard dose of 1 nmole~kg p~ntagastrin. A
compound also attenuat~s the amount of ga~tric acid WO~/19253 ~ ~ 6 7 ~ ,~fUS92~03015 secreted in response to a 1 nmole/kg dose of pentagastrin ~initial pentagastrin respons~
254 ~moles/l H+~ after the compound (cumula~ive dose of 1.1 ~mole/kg) 128 ~moles~l H+). With both compounds the antagonism is reversible with full reco~ ry of the respon~e to pentagastrin.
The compounds of the instant invention are also useful as antiulcer agents as di~cussed hereinbelow.
Aspirin induced gastric damage is a sessed in groups of 10 rat~ each.
All animals are fasted for 24 hour~ before a~d throughout the expeximent. Drug or vehic}~ i~ given 10 minutes b~fore an oral do3e of 1 ~ of a 45-mg/~L
suspension of aspirin in O.5% carboxymethylcellulo~e (CMC).
The animals are sacrificed fi~e hourq at~r aspirin administration and the ~tomach~ remo~ed a~d ope~ed ~or exami~ation.
Gastric damage wa~ scor~d as follows:
Score 1 Small hemorrhage 2 ~asge hemorxhage 3 Small ulcer 4 ~arge ulc~r ~5 5 Perforated ulcer The mean ulcQr ~core in th~ saline control group is 12.1 ~ 6.85 (tSD). Treatment with ranitidine (15 mg/kg PO) inhibits ulcer formakion by 74% gi~ing an ulcer ~core of 3.2 ~ 2.35 (p ~0.001 compared with controls). Treatment with a compound of the in~ention (10 mgtkg PO) results i~ an ulcer scor~ of 6.3 ' 4.14 (p <0.05 compar~d with controls), a 48% reduction in ulcer formation. -The ~pecific dosages employed, however, may be varied depending upon the patien~, the ~ever~ty of the :: .
WO 92/19~3 PCT/US92/03015 21067h~ :
2 4 -- ~
condition being treated, and the activity ~f the -- -compound employed. Determination of optimum dosages is within the s}cill of the art.
The cc>mpounds of khe irls~ant invention are also ~;
S u~eful as anxiolytic agent~ as described and diQcussed below .
Anxiolytic activity is asses ed in the light/dark ;
exploration test in the mou~e (B. J. Jones, et al, Br. J. Pharmacol . 93: 985 993, 1988) .
Tha nw~er of mice iq 5 and the pretreatme2~t time i~ 40 minutes. The compolaad is given p.o. in 0.1, 1, :
as~d 10 rng/lcg do~es. - ~ ..................... - ` ;
The ap~aratus is a~ op~n-topped'box, 45 cm long, 27 cm wide, and 27 cm high, di~rided into a ~mall (2/5) area and a large (3/5) area by a partil:ion that extends 20 cm above the walls . There is a ~ ~-7.5 x 7.5 c:m op~ning in the pzlrtition a~ . loor le~el. ;`~
The small compa:rtiaent is pain~:ed black and the larg~
compartment white . The floor o each co~npartmerlt is marked into 9 cm Qquares . T~e white compartment is illuminated by a 100-watt tungsten bulb 17 cm abo~e the box and the black compartment by a ~imilaxly plac~ 60--watt red bulb . The la}: oratory is .
illuminated with red ligh~.
All tests are performed between 13 hundred hours, O minutes and 18 hundred hour~, O minut~s. Each mou~e is tested by placing it in the center of the white are~ and all~wing it to explor~ the no~el en~ironment for fi~e mis~utas. Its behavior i recorded on :~
Yideotape and the beha~rioral analysis is p~rf~rmed sub~equ~ntly :Er~m the recordirlg. Fi~re parameters are measured: the latency to entry iSltO the dar3c compart~:
ment, the time spent in each area, the nus~ber of transitiorl~ between compa~m2nts, the nurl~ber of lines ,, `.~
''' `~ ~
WO9~/19~3 P~T/~S9t/03015 -25 2 ~ 0 ~ ~ 6~
crossed in each compartment, and the number of rears in each compartment.
In this test an increase in the time spent in th~
li~ht i3 a ~ensitive measure of~ that is directly related to, the anxiolytic effects of ~everal standard anxiolytic drugs. Drugs are dis~ol~ed in water or saline and administered either subcutaneou~ly, intraperitoneally, or by.mouth (PO) ~ia a stomach needle.
Compounds are active by the subcutan~ous ~oute.
Co~trol animals show 3~ cros~ings i~to th~ dark area o~ar fi~e-minut~ ~ea~urement periodQ. Mice treated with 1 mg/kg ~SC) of a compound how 85 cro3sings into the light area and only 24 crossings into the dark area, a significant (p <0.01) differenc~ from the control anxiou~ mice. Diazepam ~0.25 mg/kg IP) has an effect id~ntical to a ~ompound in the sxme ~xpsriment.
Th~ compounds of the inQt:ant in~entio~ are u~ful as ant~p~ychotic agents. Compound~ are te~t~d ~or their ability to reduc~ the effects of inkra accumbens amphetamine in ~.h~ rat as de~cribed hereinafter.
~ale Sprasue Dawley (CD) Bradford ~tr~in rats are u~ed. The rats are hous~d in groups of fi~e at a temperature of 21 2C on a 12 hour light-dark cycle '-~
of lights-on between 07 hours 00 minutes and 20 hours 00 minutes. Rat~ are fed CRM di~t (Lab ure) and all~wed water ad libitum. ~
Rats ~re anesthetized with chloral hydrate ~`:
(400 mg/kg SC~ and placed in a Kopf stereotaxic frame. ~:
Chronically indwelling guide ca~nulae (constructed of stainless ~teel tubing 0.65 ~m diamet~r held bilaterally in Per~p~ holder ) are implanted using ~:
standard stereotaxic tec~iques to terminate 3.5 mm abo~e the c~nter o* the nucleus ~ccumbens (~nt~ 9.4, Vert. 0.0, Lat. 1.6) or 5.0 mm abo~e th~ c~ntral wo 92/~92~3 2~ 6 S PCI/US92/03~t5 nucleus of th~ amygdala [Ant - 5 . 8, Vert . - 1 . 8, Lat . i:4 . 5) (atlas of ~e Grs:>ot, 1959~ . The guides are kept patent during a 14-day r~coYery period usiny :
stainless ~teel stylets, 0.3 mm diameterl which extend 0 . 5 mm beyond the guide tip~
Rats are manually r~strained and the stylets remo~ed . Intracer~3bral isl jection can~ulae, O . 3 mm diasneter, are in~erted and drugs delivered in a volume of 0 . 5 ~1 over 5 ~econds (a fu:rther 55 seconds ~a~
allowed for d~3position) ~rom Has~Lilton ~yringe~
attached via polythene tubing to the iEljec:tior~ UAitS.
~nirnals ar~ used on a single occasion os~ly. :
BehaYioral experim~rits are conducted between 07 hours 30 minutes and 21 hours 30 minutes in a quiet room maintained at 22 + 2C. :Rats are taken from the ~:
holding room and allowed one hour to adapt to ~he new environmer~t. ~ocomotor acki~ity is as~e~ed i~
indi~ridual ~creen~sd Per~pex caS~s ~5 x 15 x 15 cm `~
(high) ~banked in groups of 30) eaeh ittad with on~ pho'ocell unit along the i.~nger axis 3.5 cm ~o~
the side; this pc~sition has be~n f o~d to minimize spurious acti~ity countg due to, for example, preening ;~
and head mo~ements when the animal is stationary. ~:
Interruptio~s of the light b~am are recorded e~ery 2~ 5 minutes. At this time animal~ are al~o ob3erved for ~he pre~ence o~ any n~nspecific change in locomotor activity, ~.g., sedation, prostration, ~tereotyped -~
movements, that could interfere with the recording of -~
locomotor acti~ity.
The abilities of compounds of the in~entio~ to `~
inhibi~the hyperactiYity caused by the inj~ction of .
amphetamine into the nucleus accumbens of the rat i~
measured~
An increaQe i~ locomotor acti~ity follows the bilat~ral in~ection of amphetamine (20 ~g) into the WOg2/19253 PCT/US92/03015 -27- 21~7~
nucleus accumbens; peak hyperactivity (50 to 60 counts 5 minutes~1) occur 20 to 40 minutes after injection.
Intraperitoneal injection of tha rats with a co~pound at 20 mg~kg ~r 30 mg~kg or at 10 mgJkg reduces the hyperactivity caused by the intra-accumbens injection of amphetamine. This teQt is known to be predicti~ of antipsychotic activity ~Costall, Domeney & Naylor & Tyers, Brit~ J_Pharmac 92:881-89~).
O
- EXAMæLE 1 N-[(l,l-dimethylethoxy3car~o~yl]~-methyl~;D~-tryptophyl-~L-methionyl-L-~-a~pastyl-L-Phenylalaninamide Boc-D MeTrp-~et-~sp Phe NH2 (1) A solution of A~p-Phe-N~2 (0.6 g, 2~02 mmol) in DMF tl0 mL) was treated with cliisopropylethylamine (261 mg, 2.02 mmol~ followed by Fmoc~methioni~e ::
pentafluorophenyl ester (Fm~c~-~et-OPFP; 1.10 g, ~.
2.05 mmol) a~d the mixture stirred overnight. Removal ~.
of the D~F was followed by trituratio~ of the residue ~ -with diethyl ether:ethyl acetate (5:1) and the resulting ~olid ~iltered. This crude ~olid/
Fmoc-Met-~sp-Phe-NH2, could be u~ed without further purification ~1.22 g, 96%). ~epeated to gi~e 1.29 g :~:
of product. The solid prepared abo~e (2.00 g, 3.16 mmol3 was di~olved in DMF containing piperidine ~:~
t20~ to 50% ~/~, 16 mL) and stirred ~or 20 minutes. ~:
Remo~al of the DMF and piperidine was follow~d by `~
trituration of the residue with ether, yielding a white solid, Met-Asp-Phe-NH2, in ~uantitati~e yield.
This too could be used further without purification.
The free tripeptide (205 mg, 0.~ mmol3 was suspended in ~MF (5 mL) and diisopropylethylamine WOg2/l9253 PCT~US92/03015 21~67~j (129 mg, 1.0 mmol) added. Boc--DL MeTrp-OPFP (242 mg, ~-O.5 mmol) was added and the mixtuxe stirred until clear (approx. 2 days). Removal of ~olatiles left a light tan rasidue which was triturated with diethyl ether: ethyl acetate (1:1) giving a near white solid.
Chromatography of this solid (silica gel, 5% ~eOH in dichloromethane with 1% AcOH) yield~d a nsar white solid, 279 mg, 78% yield..
NMR (D~SO-d6 ~ 1.21 and 1.28 ~3H, 2s), 1.42 and 1.43 ~:
(9H, 2s), 1.99 and 2.01 (3H, 2s~, 2.24-2.54 (8H, m), -~
2.81 (l~, dd~, 3.11 (2H, dm), 4.18-4.4 (3H, m), 4.5 ~:-~lH, m), 6.78 (lH, br.s), 6.86-7034 ~ll~, m), 7.47 (lH, d), 7.52 and 7.41 ~lH, 2br;s),-7.7 tl~, br.m), 8.06 (lH, br.t), 8.24 (lH, br.d). :.
FAB-MS (+ve ion) 749 (M~K), 733 (~+Na), 711 ~+~
~-~e ion) 709 (~
~icroanalysiS; Calc- for C35H46N6O8S 0-75 H2O~
C, 58.03; ~, 6.54; N, 11.60; S, 4.42. -~.
Fou~d: C/ 58.13; H, 6.48; N, 11.35; S, 4~40.
~0 '.~
Similarly prepar~d was N~[t~ dimethylethoxy) ::
carbonyl~-a-methyl]-L-tryptophyl]-L-methionyl-L~
aspaxtyl-~-phenylalaninamide ~;
. ~ . . .
Boc-L-~-MeTrp-Net-Asp-Phe-N~2, (2), and N-[(l-1-dimekhyl~thoxy)carbonyl]-~-methyl-D- :
tryptophyl-L-methionyl-L-a-aspartyl-L~
phenylalaninamide Boc-D-~-~eTrp Met-Asp-Phe-NH2, (3). ~
~ ;
NMR (2; D~SO-d6) ~ 1.25 (3H, s), 1.44 ~9H, s), 1.83 ~2H, m), 2.01 (3H, s~, 2.38 (2H, m), 2.65 ~lH, m), :~
2.82 (lH, m), 4.20 (lH, m)~ 4.33 (lH, m), 4.50 (lH, W~92/19253 PCT/US92/03015 -29 21~76~
m), 6.92-7.47 tl2H, m), 7.82 (2H, m~ r 8 . ll (lR~ br.d), 10 . 91 ~l~I, s) .
.
N2~ ~3; DMSO--d6) ~ 1.29 (3H~ s) r 1.44 ~9H~ s) ~ 1.92 (2Hr m), 2.02 (3Hr s) r 2 . 4~ (2H~ t) ~ 2 . 60 ~ td) r 2.84 (lHr dd) t 3.13 (2H~ d~ r 3.31 (. H+water), 4~19 (l~r m), 4.36 (lH, m), 4.48 (lH, m), 6.94-7.24 (12Hr m) r 7.32 (lHr d) ~ 7.47 (lX~ d) r 7.84 (lHr br.d~, 8.14 (l~, br.d~, 8.22 (1~, br.s), 10.92 (lH, s).
EX~NPL~ 2 Also prepared by this method were a-me~hyl L ,.
tr~plophyl ~-methionyl L-a~a~partyl~
phenylalaninamide :
L-a-~eTrp-Met-Asp-Ph~;NH2, (4~
:. ' ' a-methyl-D-tr~ptophyl-L~methio~yl-L-a-aspartyl-L-phenylalaninamide D~a-MeTrp-Met-Asp-Phs-NH2, (5) and ''';:
~-methyl~DL-tryptophyl L-methionyl-L a-aspartyl~L
phenylalanin~mide ~
DL-~-MeTrp-Met Asp Phe-NH2, (6). ~-In these latter three instances the final residue used was the appropriate E'moc-a-MeTrp activated ester 3~ leading to the protected tetrapeptide which was ~;
treate~. with piperidine in DMF to remove the . :~
pxotecting group. This furnished the completely deprotected tetrapeptides (4-6). Purificatiosl was by RP~HPLC (MeCN-H20/0.1~6 TFA) yielding the trifluoroacetate salt of each peptide.
WO 92/1~253 P~/U~g2/03015 2 1~ 67 ~ -30-N~ ~4 ; D203 ~ 1 . 73 (3H, s), 1 . 85 (2H, q), 2 . 04 ~3.H, s), 2 . 34 (2H, t), 2 . 66 (2H, qd), 3 . 07 ~2H, ~d), 3 . 41 (2~, s), 4 . 34 (lH, t), 4 . 54 (2H, m), 7 . 14 (lE~, t), 7 . 21-7 . 37 (9EI, m), 7 . 48-7 . 56 (2El, dd) .
N~ ~5; D20) ~ 1. 54 (3EI, s), 1. 713 (2~I, m), 1 . 97 (3H, s), 2.54 (2~I, m), 2.93-3.28 (6H, m), 4.13 (l}I, ~), . 42 (lX, t), 4 . 55 ~1~, m), 7 . ~ 3--7 . 36 (8H, m), 7 . 51 ~.
(l.H, d), 8.18 ~ d). .`;
N~ (6; D20) ~ 1.73 (3H, ~) j 1.85 (2~, m), 1.94 ~d 2 . 03 (3HI 2s), 2 . 34 (lH, t), 2 . 74 ~2H, m), 3 . 06 (2 qd) ~ 3.40 (2H~ abq) ~ 4 . 01 a~ld 4 .34 (lH~ 2t:) r 4. 60 (2H~
m), 7.11-7.48 (lOH, m), 7~53 (2~, t). ~
~-The following compounds were made by the proces e~
described above for ~ethod A. -~-Glycyl-amethyl-D-tryptophyl L~methionyl-L-a-a~partyl-L-phenylalaninamide ~-"
~Gly-DMeTrp-~et-~p-Phe-NH.2), Glycyl-a-methyl DL tryp~ophyl-L-methi~nyl~
aspartyl~L-phenylalaninamide (Gly-DL~eTrp-M~t-A~p-Phe-N~23, Glycyl~a-methyl~-tryptophyl-L-methionyl-~ a~
aspartyl-L-phenylala~inamid~
(Gly--L~EeTrp--~et Asp--Phe--N~2), T~ et -A~p-D~-MePhe--NH2 t::F3C02H, and Trp--Met-DL--~qeA.sp--Phe NH2- CF3C02H.
33 They c:an also be made by the following example.
. ........................................................... :, W~92/19253 PCT/US92~03015 -31- 21 ~7~
EXAMP.LE 3 -~ryptophyl-a-methyl-DL-methionyl-L~a-aspartyl-k-phenylalaninamide ~-Trp--DL-~-M~Met--~sp-Phe--NH2 (7) Using a No~a Biochem 4170 automated peptide synthesi~er and Bioplus~ software the peptide was constructed from the C-terminus using Fmoc amino acid pentafluorophenyl or DEBt esters and HOBt cataly~is.
Each rssi~ue was pre~ent in a fi~efold ~xcess to en3ure rapid and complete aeylation. O~ a 0.095 mmol scale, the peptide (4) was i801ated ~llo~ing TFA
clea~age (94% ~F~, 5% ani~ole, 1% ~anedithiol) from :~
the re~in (2h, room ~emperature) in 46~ yield (32 m~
isolated). ~PLC (ClB, 20-80% MeCN-H2O ~ 0.1~ TFA3 sho~ed greater ~han 90% puri~.y ~254 ~m detection) as a 1:1 mixture of dia~tereomers.
N~R (D2O) ~ 1.03 ~3H~ s), 1.69 ~2~, m), lv91 ~2H, m), 1.95 ~3~, s~, ~.5g (2H, m), 3.09 (2~, m), 3.38 ~@H, ~:~
m), 4.31 ~lH, m~, 4.51 (2H, mJ r 7.17 - 7.37 (10~ m), 7.52 (lH, d~, 7.6B ~lH, d), 10.94 (lH, 5). ~hen produced by solution-pha3e methods the product was :;~
iderltiral. ~' Similarly prepared was I. ~-M[eTrp ~let~Asp--Phe N~2, (4), identical is~ eYery r~pect to that prepared abo~e.
EX~I,E 4 :.
Trp--~et~sp--DL--MePhe N~I2 CF3C02H (8~ ~
Fmoc: ASp ( OBut )--DL--MePhe--N~2 , ~ . :
A mixture of MePhe NH2 (90 mg, 0.5 mmol) and Fmoc-Asp (OBu- ) OP:Ep ( ~ 90 mg, O . 5 mmol) were dissolved in the minimum qu~Lntity of ethyl acetate and the solvent WO ~2/~9253 PCr/U~92/03~15 2,10676i 32 immedii3tely distilled away. Th~ re~ulting viscous oil was heated in a boiling water bath for 2 hours, then cooled. The resulting resin was triturated with diethyl ether from which the product precipitated.
Yield 159 mg (56%) ~ further crop o~ product could :
be obtained by chromatography of the ethereal washinys .
.'-.
F;noc-~5et--A .p (OESut~ -DL--~ePhe-NE2 The preceding dipeptide (9V0 mgr 1.58 mmol) was N--deprotscted using 2 0% piperidine in D~F ~or 2 0 minutes then the ~olvant removed completely . Th~
resic~ue was repeatedly triturat~d with s~ hexane.
lea~ing the fres amir.o dipeptide (293 mg, 0. 84 mmol~
To this was add~d Fmoc-~5et OPfp (450 mg, 0 . 84 mmol) :
f ollowed by the minimum ~uantity of ethyl ac~tate to gi~e a clear solutio~. The ~olve~t was immediately -:
removed and the resulting ~i~cou~ oil heated in a water bath at 65~C for 45 minut~s. Trituration of l:h~
2 0 resulting re3in with diethyl ether yielded the .
protect~3d tripeptide (418 mg, 78%).
Fmoc-Trp~Met Asp (OBut)--DL MePh~--N~2 The preGedins~ tripeptide ~300 mg, 0 . 43 mxnol) ~as N-deprotected u~in~ 20% piperidine in DME? ~or 20 minutes. ~emo~ral of the ~olvent followed by ~rituration o~ the residu~ with n-hexane le~t th~ free tripeptide (198 mg, 0 . 41 mmol) . To this was a ded ~moc-Trp~OPfp ~243 mg, 0 . 41 mmol) and sufficient ethyl acetate to gi-re a clear solution. The solvent wa~
immedia~ely remo~ed and the result~ ng mixture heated at 654C for 15 minutes. Trituration vf the residue with diethyl ether ga~re a white solid, 283 mg, 7896.
WO 92/192~3 PCr/l~S~2/030lS
210~7~3 Trp-Met-Asp-DL-MePhe NH2 CF3t::02H ( B ) .
The preceding tetrap~ptide was M-deprotected using 2û% piperidine in D~F for 20 minutes then the solvent ~:
remo~ed and the residue triturated with n-h~xane, lea~ring the fr~e amino tetrapeptide. This was treated ~-with thic~anisole and ethanedithiol (100-250 ,UL~mmol and 50-100 ~IL/mmol) followed by 95% aqueous TFA
(3-5 mL/mmol) for 5 minutes. All volatiles were remo~red and the residue triturated wikh diethyl ether, gi~ving a white solid, puri~ied by pr~parati~re ~ C.
Trp--~qet--DL--MeAsp--Phe-NH2 CP'3C02H (9) Fmoc-DL--MeAsp (OBut)--Phe--NH2 Fmoc--DL-MeAsp (0}3ut) 0Pfp ~1. 46 g, 2 . 47 mmol) and H-Phe-NH2 (O . 4~ g, 2 . 5 mmol) were dissolved in the :
mini~num quantity of ethyl acetate to gi~e a cl~ar solution~ then the ~301~rent ~s immediately res~oY~d.
The mixtur~ wa~ then heat~d at 100C for 1 hour and .-cool~d . Tritur~tion of the residue with diethyl ether ~ ~
yielded th protect~d dipepti.de, 993 mg, 71%. ~:.
Fmoc-~et-DI.-MeAsp (OBut)--Phe--NH2 .:.
The preceding dip~ptide (571 mg, 1. 0 mmol) was N-deprot~cted as pre~iou ly describad. Following trituration with n-hexane, the residue was mix~d with Fmoc-~et--OPfp (350 mgl 0 . 65 mmol) ar~d suffi ::ierlt ethyl ~ `:
acetate to dissol~re. Immediate removal of the ~ol~rent ~.
was followed by heatirlg the residue at 65C for 4 hours. Trituration of the residue with diethyl ether....~rielded the protected tripeptide, 210 mg, 4696.
A further crop could be isolated from the ethereal washinys by chromatography.
.... ... .. ..
WO 92/192~3 PCrfUS92/03015 2 1 ~ ~ 7 6~ ~34~ ` ~ ~
Fmoc-Trp-Met -Dl,-MeA~p ( OBut )--Ph-NH2 The above tripeptide (400 mg, 0.57 mmol) was N--deprotected as pre~riously described. Following trituration of the re~idue with n-hexane, 258 mg of ~-:
product were i~;olated. 200 mg (0.4? mmol) of this and Fmoc-Trp-OP:Ep (246 mg, 0 . 42 ~unol) ~as added ~ith ~ -sufficient ethyl acetate to dissol~e. The solverlt ~as immediately remo~ed and the residue heated at 65C fox .
2 0 minu es . Trituration of the resulting mixture with l 0 diethyl ether yielded the protected tetrapeptide, ~ -283 mg, 7796 . .
.
Trp-Metol~ Me~p-Ph~-NH2 CF3C02~ ( 9 ) The preceding tetrapeptide was treated as the tetrapeptide, yielding (8~ and gave after final trituration with ether.
.. ~
CCK8s Agonist 1~45 . 4 :l --~''""' , The compounds were further provén to be agonists following experaments in which they ~exe challeng~d with ~he previ~usily de~cri~ed ~otent and selecti~e CCK-B antagonist, reference compound PD 134308 named ~R-(R*,R*)-4~l2-~3-(lH-indol-3-yl)-2-methyl~1-oxo-2 ~[~tricyclo r 3 ~ 3 .1.13-7~ dec-2-yloxy~carbonyl]amino~
propyl]aminoJ-l-phenethyl]amino~-4-oxobutanoic acid.
TABLE IV
C:ompound Graph(PD 134308) /s~M
(4) + ref~rence compour~d 44 . 67 (5) + refçrenc:e compound 43 ~ 02 ~6) ~ reference compouslcl 3 14 . 4 CCK8~ ~ refererlce compound -- 5 . 4 Data shows ~hat compound (4) is equipotent with CC~8s while compound (6) possesses slightly lower el~ctrophysiological activity and compound (53 has relatively low agonist acti~ity. Compound ~2) îs a partial agonist.
WO~2/192S3 P~T/VS92/03015 ~13~ 76j -20~
Compounds of the present invention are useful as appetite suppre~a~ts as based on the tests described hexei~below.
I~ the Palatable Diet Feeding a~say, ~dult male Hooded Lister rats weighing between 200-400 g are hou~d indi~idually and trained to ~at a palatable diet. This diet consists of Nestl~s sweetened condensed milk, powdered rat food, and rat water which was when blended together set to a firm consi tency.
Each rat is pre~Qnted with 20 to 30 g of the pala~bl~
- diet for 30 minutes per day during the light pha~e of the lght-dark cycle o~r a trai~ing period o~ S day~
The inta~ ~f palatable diet is mea~ured by weighing -~
the food container before and after the 30~minute `~
access period (limits of ac~.uracy O.l g). Care is taken to collect and correct for a~y spillage of the diet. R~t~ ha~e free acces~ to pellet food and water ~;
exc~pt during the 30-minut~ t~t period. ~
After the traini~g perivd, do e~re~ponQ~ ~urve~ ::
are construct~d ~or CCX8 and several repre~ntati~e compounds of the present in~ention (n - 8-lO xats per do~e le~el). ~PE50 ~a~ue~ ('95% confidence limits) are obtained for the anorectic effeats of th~3e compounds.
In thera~eutic u~e as appetite Quppression :~
ag~nts, the compounds of the in tant invention are admi~istered to the patient at dosage le~els o from about 200 to about 2800 mg per day. . ~.
As ago~ists the compounds of the instant in~ention ha~e therapeutic uti~ity as appetite suppr~ants and~ as antagonists, as agents in ~.
treating ga~trointestinal disorders, central ner~ous ~ystem di~order~ such as CNS suppressants which can ;~
exhibit such effect~ as antipsychotics, n~uroleptics, anxiolytics, and antico~ulsants.
WO 92/lg2S3 PCrJUS92/~3015 210~7&~
As antagonists, the compounds o~ the instant invention are expected to have utility in reducing gastric acid secretion, in raducing anxiety, in : - ~
treating gastrointestinal ulcers , in treating ~:
psychotic behavior, in blockis~g the reaction caused by withdrawal from drug, alcohol, cocain~3, benzodiazepine, diazepam, or nicotine, and in pot~3ntiating the e:Efects of morphine for treating pain .
CCK A~alo~s c:ontaining o~ znethyl amiIlo acid~ were mad~ by three me~hods. ~ethocl A is a solutioxl phase procedure ~herein the peptides were a~bled uging standard 301Util:~n pha~e protocols yielding ~u~ ly protected interrnediates or standard salt coupling which yield a~partic acid side-chain d~protected intermediates. Examples 1 and 2 below illustrate Method A.
Th~3 peptides c:a~ also be prepared by ~ethod B, a solid-pha~ method wherein peptid~3~ were con. tructed on solid~pha^~ rg~inc d ~igned to produc~ C-termirlal amides either by tre~tment of the resin with a~unonia in methanol s~r by direct clea~age of an appropriate~ y ~:ubstituted resin usin~ trifluoroacetic acid, with the reguired ~c~engerr, gi~ing the amides directly. The latter protoGol was u ad ~ith DuPont ~apidAmide~ or Nova Biochem No~a~yn KR~ resins either in a simple bubbler apparatus ~DuPo~t re~in) or automated synthesizer (Nova Biochem re~in). This method is illustrated by Example 3 b~low.
The third method, ~ethod C, is a non~ol~t method whereby the amino compo~e~t ~nd a pentafluoroph~yl e~ter ar~ heated together without a solvent at 50~ to 100C for lO to 120 minutes or more.
ThiQ method is illu~trated by E~ampl~ 4 b~low.
~ .
WO 92/192S3 PCr/US92J03015 21 1~ ~7 ~3 Male Hooded ~ister rats (175--2~0 g) are housed individually and fasted overnight (free accese to ` ~:
watex~ . They are arlesthetized with urethane ~1. 5 g/ky IP) and the trachea ca~ulated to aid ~pontaneous `:
respiration. The stomach i~ perfused conl;inuously using a modification of the original method of Ghosh & ~ ~
Schild in "Continuous recording of acid secretion in . ~' the rat", Br._J~aE~. 13:54--61, 1956 as described :~:
by Parsons in "Quantitative studi~3s of drug-inducecl gastric acid ~ecretion". ~Ph.D. Thesis, Uni~ersity of London, 1969? . Th~ ca~ity of the: ~tomach is per~u ed at a rate of 3 ~/min with 5.4% w~v gllleose ~olution ~:
throu~h both the e~phaç~eal and body cannula. ~he fluid is propelled by a roll~3r pump (Gilson, ~inipuls 2), through heating coils to bring its temperature to 37 ~t 1C. The perfusion ~luid i~
collected by the fundic collecting f~nnel and pas~ed to a p~ electrode connected to a Jenway pH met2r ~PE~6~. ~n output is taken from the p~ meter to a Rikadenki chart recorder f or the on~line recording of the p~ of the gastric perfu~ate.
Pentagastrin is stored as a frozen aliquot and diluted to the r~quired concentrations ~ith st~rile O . 9% w/~r NaCl . No~rel com~c~unds are diæ~3~1~ed in :
steril~ 0. 9% w/~r NaCl on the day of the experiment. ~:
Drugs are administered I'V through a canslulated jugular :
~rein s a bolus in a dose ~olume of 1 mL jkg wa~h~3d in with 0 .15 m~ O . 9% w/~ NaCl. Ba~al pH is allo~ed to ~`.
stabilize ~ef or~ administration o~ compourld~ is begun .
Typically 30 minutes elap~3es between surgery and the :Eirst .~ompound a~ninistration . ~:
Compounds of the in~rention may al~o ar~tagonize the stimulation of 5~astric ac:id secretion produt::ed by ~;~
a standard dose of 1 nmole~kg p~ntagastrin. A
compound also attenuat~s the amount of ga~tric acid WO~/19253 ~ ~ 6 7 ~ ,~fUS92~03015 secreted in response to a 1 nmole/kg dose of pentagastrin ~initial pentagastrin respons~
254 ~moles/l H+~ after the compound (cumula~ive dose of 1.1 ~mole/kg) 128 ~moles~l H+). With both compounds the antagonism is reversible with full reco~ ry of the respon~e to pentagastrin.
The compounds of the instant invention are also useful as antiulcer agents as di~cussed hereinbelow.
Aspirin induced gastric damage is a sessed in groups of 10 rat~ each.
All animals are fasted for 24 hour~ before a~d throughout the expeximent. Drug or vehic}~ i~ given 10 minutes b~fore an oral do3e of 1 ~ of a 45-mg/~L
suspension of aspirin in O.5% carboxymethylcellulo~e (CMC).
The animals are sacrificed fi~e hourq at~r aspirin administration and the ~tomach~ remo~ed a~d ope~ed ~or exami~ation.
Gastric damage wa~ scor~d as follows:
Score 1 Small hemorrhage 2 ~asge hemorxhage 3 Small ulcer 4 ~arge ulc~r ~5 5 Perforated ulcer The mean ulcQr ~core in th~ saline control group is 12.1 ~ 6.85 (tSD). Treatment with ranitidine (15 mg/kg PO) inhibits ulcer formakion by 74% gi~ing an ulcer ~core of 3.2 ~ 2.35 (p ~0.001 compared with controls). Treatment with a compound of the in~ention (10 mgtkg PO) results i~ an ulcer scor~ of 6.3 ' 4.14 (p <0.05 compar~d with controls), a 48% reduction in ulcer formation. -The ~pecific dosages employed, however, may be varied depending upon the patien~, the ~ever~ty of the :: .
WO 92/19~3 PCT/US92/03015 21067h~ :
2 4 -- ~
condition being treated, and the activity ~f the -- -compound employed. Determination of optimum dosages is within the s}cill of the art.
The cc>mpounds of khe irls~ant invention are also ~;
S u~eful as anxiolytic agent~ as described and diQcussed below .
Anxiolytic activity is asses ed in the light/dark ;
exploration test in the mou~e (B. J. Jones, et al, Br. J. Pharmacol . 93: 985 993, 1988) .
Tha nw~er of mice iq 5 and the pretreatme2~t time i~ 40 minutes. The compolaad is given p.o. in 0.1, 1, :
as~d 10 rng/lcg do~es. - ~ ..................... - ` ;
The ap~aratus is a~ op~n-topped'box, 45 cm long, 27 cm wide, and 27 cm high, di~rided into a ~mall (2/5) area and a large (3/5) area by a partil:ion that extends 20 cm above the walls . There is a ~ ~-7.5 x 7.5 c:m op~ning in the pzlrtition a~ . loor le~el. ;`~
The small compa:rtiaent is pain~:ed black and the larg~
compartment white . The floor o each co~npartmerlt is marked into 9 cm Qquares . T~e white compartment is illuminated by a 100-watt tungsten bulb 17 cm abo~e the box and the black compartment by a ~imilaxly plac~ 60--watt red bulb . The la}: oratory is .
illuminated with red ligh~.
All tests are performed between 13 hundred hours, O minutes and 18 hundred hour~, O minut~s. Each mou~e is tested by placing it in the center of the white are~ and all~wing it to explor~ the no~el en~ironment for fi~e mis~utas. Its behavior i recorded on :~
Yideotape and the beha~rioral analysis is p~rf~rmed sub~equ~ntly :Er~m the recordirlg. Fi~re parameters are measured: the latency to entry iSltO the dar3c compart~:
ment, the time spent in each area, the nus~ber of transitiorl~ between compa~m2nts, the nurl~ber of lines ,, `.~
''' `~ ~
WO9~/19~3 P~T/~S9t/03015 -25 2 ~ 0 ~ ~ 6~
crossed in each compartment, and the number of rears in each compartment.
In this test an increase in the time spent in th~
li~ht i3 a ~ensitive measure of~ that is directly related to, the anxiolytic effects of ~everal standard anxiolytic drugs. Drugs are dis~ol~ed in water or saline and administered either subcutaneou~ly, intraperitoneally, or by.mouth (PO) ~ia a stomach needle.
Compounds are active by the subcutan~ous ~oute.
Co~trol animals show 3~ cros~ings i~to th~ dark area o~ar fi~e-minut~ ~ea~urement periodQ. Mice treated with 1 mg/kg ~SC) of a compound how 85 cro3sings into the light area and only 24 crossings into the dark area, a significant (p <0.01) differenc~ from the control anxiou~ mice. Diazepam ~0.25 mg/kg IP) has an effect id~ntical to a ~ompound in the sxme ~xpsriment.
Th~ compounds of the inQt:ant in~entio~ are u~ful as ant~p~ychotic agents. Compound~ are te~t~d ~or their ability to reduc~ the effects of inkra accumbens amphetamine in ~.h~ rat as de~cribed hereinafter.
~ale Sprasue Dawley (CD) Bradford ~tr~in rats are u~ed. The rats are hous~d in groups of fi~e at a temperature of 21 2C on a 12 hour light-dark cycle '-~
of lights-on between 07 hours 00 minutes and 20 hours 00 minutes. Rat~ are fed CRM di~t (Lab ure) and all~wed water ad libitum. ~
Rats ~re anesthetized with chloral hydrate ~`:
(400 mg/kg SC~ and placed in a Kopf stereotaxic frame. ~:
Chronically indwelling guide ca~nulae (constructed of stainless ~teel tubing 0.65 ~m diamet~r held bilaterally in Per~p~ holder ) are implanted using ~:
standard stereotaxic tec~iques to terminate 3.5 mm abo~e the c~nter o* the nucleus ~ccumbens (~nt~ 9.4, Vert. 0.0, Lat. 1.6) or 5.0 mm abo~e th~ c~ntral wo 92/~92~3 2~ 6 S PCI/US92/03~t5 nucleus of th~ amygdala [Ant - 5 . 8, Vert . - 1 . 8, Lat . i:4 . 5) (atlas of ~e Grs:>ot, 1959~ . The guides are kept patent during a 14-day r~coYery period usiny :
stainless ~teel stylets, 0.3 mm diameterl which extend 0 . 5 mm beyond the guide tip~
Rats are manually r~strained and the stylets remo~ed . Intracer~3bral isl jection can~ulae, O . 3 mm diasneter, are in~erted and drugs delivered in a volume of 0 . 5 ~1 over 5 ~econds (a fu:rther 55 seconds ~a~
allowed for d~3position) ~rom Has~Lilton ~yringe~
attached via polythene tubing to the iEljec:tior~ UAitS.
~nirnals ar~ used on a single occasion os~ly. :
BehaYioral experim~rits are conducted between 07 hours 30 minutes and 21 hours 30 minutes in a quiet room maintained at 22 + 2C. :Rats are taken from the ~:
holding room and allowed one hour to adapt to ~he new environmer~t. ~ocomotor acki~ity is as~e~ed i~
indi~ridual ~creen~sd Per~pex caS~s ~5 x 15 x 15 cm `~
(high) ~banked in groups of 30) eaeh ittad with on~ pho'ocell unit along the i.~nger axis 3.5 cm ~o~
the side; this pc~sition has be~n f o~d to minimize spurious acti~ity countg due to, for example, preening ;~
and head mo~ements when the animal is stationary. ~:
Interruptio~s of the light b~am are recorded e~ery 2~ 5 minutes. At this time animal~ are al~o ob3erved for ~he pre~ence o~ any n~nspecific change in locomotor activity, ~.g., sedation, prostration, ~tereotyped -~
movements, that could interfere with the recording of -~
locomotor acti~ity.
The abilities of compounds of the in~entio~ to `~
inhibi~the hyperactiYity caused by the inj~ction of .
amphetamine into the nucleus accumbens of the rat i~
measured~
An increaQe i~ locomotor acti~ity follows the bilat~ral in~ection of amphetamine (20 ~g) into the WOg2/19253 PCT/US92/03015 -27- 21~7~
nucleus accumbens; peak hyperactivity (50 to 60 counts 5 minutes~1) occur 20 to 40 minutes after injection.
Intraperitoneal injection of tha rats with a co~pound at 20 mg~kg ~r 30 mg~kg or at 10 mgJkg reduces the hyperactivity caused by the intra-accumbens injection of amphetamine. This teQt is known to be predicti~ of antipsychotic activity ~Costall, Domeney & Naylor & Tyers, Brit~ J_Pharmac 92:881-89~).
O
- EXAMæLE 1 N-[(l,l-dimethylethoxy3car~o~yl]~-methyl~;D~-tryptophyl-~L-methionyl-L-~-a~pastyl-L-Phenylalaninamide Boc-D MeTrp-~et-~sp Phe NH2 (1) A solution of A~p-Phe-N~2 (0.6 g, 2~02 mmol) in DMF tl0 mL) was treated with cliisopropylethylamine (261 mg, 2.02 mmol~ followed by Fmoc~methioni~e ::
pentafluorophenyl ester (Fm~c~-~et-OPFP; 1.10 g, ~.
2.05 mmol) a~d the mixture stirred overnight. Removal ~.
of the D~F was followed by trituratio~ of the residue ~ -with diethyl ether:ethyl acetate (5:1) and the resulting ~olid ~iltered. This crude ~olid/
Fmoc-Met-~sp-Phe-NH2, could be u~ed without further purification ~1.22 g, 96%). ~epeated to gi~e 1.29 g :~:
of product. The solid prepared abo~e (2.00 g, 3.16 mmol3 was di~olved in DMF containing piperidine ~:~
t20~ to 50% ~/~, 16 mL) and stirred ~or 20 minutes. ~:
Remo~al of the DMF and piperidine was follow~d by `~
trituration of the residue with ether, yielding a white solid, Met-Asp-Phe-NH2, in ~uantitati~e yield.
This too could be used further without purification.
The free tripeptide (205 mg, 0.~ mmol3 was suspended in ~MF (5 mL) and diisopropylethylamine WOg2/l9253 PCT~US92/03015 21~67~j (129 mg, 1.0 mmol) added. Boc--DL MeTrp-OPFP (242 mg, ~-O.5 mmol) was added and the mixtuxe stirred until clear (approx. 2 days). Removal of ~olatiles left a light tan rasidue which was triturated with diethyl ether: ethyl acetate (1:1) giving a near white solid.
Chromatography of this solid (silica gel, 5% ~eOH in dichloromethane with 1% AcOH) yield~d a nsar white solid, 279 mg, 78% yield..
NMR (D~SO-d6 ~ 1.21 and 1.28 ~3H, 2s), 1.42 and 1.43 ~:
(9H, 2s), 1.99 and 2.01 (3H, 2s~, 2.24-2.54 (8H, m), -~
2.81 (l~, dd~, 3.11 (2H, dm), 4.18-4.4 (3H, m), 4.5 ~:-~lH, m), 6.78 (lH, br.s), 6.86-7034 ~ll~, m), 7.47 (lH, d), 7.52 and 7.41 ~lH, 2br;s),-7.7 tl~, br.m), 8.06 (lH, br.t), 8.24 (lH, br.d). :.
FAB-MS (+ve ion) 749 (M~K), 733 (~+Na), 711 ~+~
~-~e ion) 709 (~
~icroanalysiS; Calc- for C35H46N6O8S 0-75 H2O~
C, 58.03; ~, 6.54; N, 11.60; S, 4.42. -~.
Fou~d: C/ 58.13; H, 6.48; N, 11.35; S, 4~40.
~0 '.~
Similarly prepar~d was N~[t~ dimethylethoxy) ::
carbonyl~-a-methyl]-L-tryptophyl]-L-methionyl-L~
aspaxtyl-~-phenylalaninamide ~;
. ~ . . .
Boc-L-~-MeTrp-Net-Asp-Phe-N~2, (2), and N-[(l-1-dimekhyl~thoxy)carbonyl]-~-methyl-D- :
tryptophyl-L-methionyl-L-a-aspartyl-L~
phenylalaninamide Boc-D-~-~eTrp Met-Asp-Phe-NH2, (3). ~
~ ;
NMR (2; D~SO-d6) ~ 1.25 (3H, s), 1.44 ~9H, s), 1.83 ~2H, m), 2.01 (3H, s~, 2.38 (2H, m), 2.65 ~lH, m), :~
2.82 (lH, m), 4.20 (lH, m)~ 4.33 (lH, m), 4.50 (lH, W~92/19253 PCT/US92/03015 -29 21~76~
m), 6.92-7.47 tl2H, m), 7.82 (2H, m~ r 8 . ll (lR~ br.d), 10 . 91 ~l~I, s) .
.
N2~ ~3; DMSO--d6) ~ 1.29 (3H~ s) r 1.44 ~9H~ s) ~ 1.92 (2Hr m), 2.02 (3Hr s) r 2 . 4~ (2H~ t) ~ 2 . 60 ~ td) r 2.84 (lHr dd) t 3.13 (2H~ d~ r 3.31 (. H+water), 4~19 (l~r m), 4.36 (lH, m), 4.48 (lH, m), 6.94-7.24 (12Hr m) r 7.32 (lHr d) ~ 7.47 (lX~ d) r 7.84 (lHr br.d~, 8.14 (l~, br.d~, 8.22 (1~, br.s), 10.92 (lH, s).
EX~NPL~ 2 Also prepared by this method were a-me~hyl L ,.
tr~plophyl ~-methionyl L-a~a~partyl~
phenylalaninamide :
L-a-~eTrp-Met-Asp-Ph~;NH2, (4~
:. ' ' a-methyl-D-tr~ptophyl-L~methio~yl-L-a-aspartyl-L-phenylalaninamide D~a-MeTrp-Met-Asp-Phs-NH2, (5) and ''';:
~-methyl~DL-tryptophyl L-methionyl-L a-aspartyl~L
phenylalanin~mide ~
DL-~-MeTrp-Met Asp Phe-NH2, (6). ~-In these latter three instances the final residue used was the appropriate E'moc-a-MeTrp activated ester 3~ leading to the protected tetrapeptide which was ~;
treate~. with piperidine in DMF to remove the . :~
pxotecting group. This furnished the completely deprotected tetrapeptides (4-6). Purificatiosl was by RP~HPLC (MeCN-H20/0.1~6 TFA) yielding the trifluoroacetate salt of each peptide.
WO 92/1~253 P~/U~g2/03015 2 1~ 67 ~ -30-N~ ~4 ; D203 ~ 1 . 73 (3H, s), 1 . 85 (2H, q), 2 . 04 ~3.H, s), 2 . 34 (2H, t), 2 . 66 (2H, qd), 3 . 07 ~2H, ~d), 3 . 41 (2~, s), 4 . 34 (lH, t), 4 . 54 (2H, m), 7 . 14 (lE~, t), 7 . 21-7 . 37 (9EI, m), 7 . 48-7 . 56 (2El, dd) .
N~ ~5; D20) ~ 1. 54 (3EI, s), 1. 713 (2~I, m), 1 . 97 (3H, s), 2.54 (2~I, m), 2.93-3.28 (6H, m), 4.13 (l}I, ~), . 42 (lX, t), 4 . 55 ~1~, m), 7 . ~ 3--7 . 36 (8H, m), 7 . 51 ~.
(l.H, d), 8.18 ~ d). .`;
N~ (6; D20) ~ 1.73 (3H, ~) j 1.85 (2~, m), 1.94 ~d 2 . 03 (3HI 2s), 2 . 34 (lH, t), 2 . 74 ~2H, m), 3 . 06 (2 qd) ~ 3.40 (2H~ abq) ~ 4 . 01 a~ld 4 .34 (lH~ 2t:) r 4. 60 (2H~
m), 7.11-7.48 (lOH, m), 7~53 (2~, t). ~
~-The following compounds were made by the proces e~
described above for ~ethod A. -~-Glycyl-amethyl-D-tryptophyl L~methionyl-L-a-a~partyl-L-phenylalaninamide ~-"
~Gly-DMeTrp-~et-~p-Phe-NH.2), Glycyl-a-methyl DL tryp~ophyl-L-methi~nyl~
aspartyl~L-phenylalaninamide (Gly-DL~eTrp-M~t-A~p-Phe-N~23, Glycyl~a-methyl~-tryptophyl-L-methionyl-~ a~
aspartyl-L-phenylala~inamid~
(Gly--L~EeTrp--~et Asp--Phe--N~2), T~ et -A~p-D~-MePhe--NH2 t::F3C02H, and Trp--Met-DL--~qeA.sp--Phe NH2- CF3C02H.
33 They c:an also be made by the following example.
. ........................................................... :, W~92/19253 PCT/US92~03015 -31- 21 ~7~
EXAMP.LE 3 -~ryptophyl-a-methyl-DL-methionyl-L~a-aspartyl-k-phenylalaninamide ~-Trp--DL-~-M~Met--~sp-Phe--NH2 (7) Using a No~a Biochem 4170 automated peptide synthesi~er and Bioplus~ software the peptide was constructed from the C-terminus using Fmoc amino acid pentafluorophenyl or DEBt esters and HOBt cataly~is.
Each rssi~ue was pre~ent in a fi~efold ~xcess to en3ure rapid and complete aeylation. O~ a 0.095 mmol scale, the peptide (4) was i801ated ~llo~ing TFA
clea~age (94% ~F~, 5% ani~ole, 1% ~anedithiol) from :~
the re~in (2h, room ~emperature) in 46~ yield (32 m~
isolated). ~PLC (ClB, 20-80% MeCN-H2O ~ 0.1~ TFA3 sho~ed greater ~han 90% puri~.y ~254 ~m detection) as a 1:1 mixture of dia~tereomers.
N~R (D2O) ~ 1.03 ~3H~ s), 1.69 ~2~, m), lv91 ~2H, m), 1.95 ~3~, s~, ~.5g (2H, m), 3.09 (2~, m), 3.38 ~@H, ~:~
m), 4.31 ~lH, m~, 4.51 (2H, mJ r 7.17 - 7.37 (10~ m), 7.52 (lH, d~, 7.6B ~lH, d), 10.94 (lH, 5). ~hen produced by solution-pha3e methods the product was :;~
iderltiral. ~' Similarly prepared was I. ~-M[eTrp ~let~Asp--Phe N~2, (4), identical is~ eYery r~pect to that prepared abo~e.
EX~I,E 4 :.
Trp--~et~sp--DL--MePhe N~I2 CF3C02H (8~ ~
Fmoc: ASp ( OBut )--DL--MePhe--N~2 , ~ . :
A mixture of MePhe NH2 (90 mg, 0.5 mmol) and Fmoc-Asp (OBu- ) OP:Ep ( ~ 90 mg, O . 5 mmol) were dissolved in the minimum qu~Lntity of ethyl acetate and the solvent WO ~2/~9253 PCr/U~92/03~15 2,10676i 32 immedii3tely distilled away. Th~ re~ulting viscous oil was heated in a boiling water bath for 2 hours, then cooled. The resulting resin was triturated with diethyl ether from which the product precipitated.
Yield 159 mg (56%) ~ further crop o~ product could :
be obtained by chromatography of the ethereal washinys .
.'-.
F;noc-~5et--A .p (OESut~ -DL--~ePhe-NE2 The preceding dipeptide (9V0 mgr 1.58 mmol) was N--deprotscted using 2 0% piperidine in D~F ~or 2 0 minutes then the ~olvant removed completely . Th~
resic~ue was repeatedly triturat~d with s~ hexane.
lea~ing the fres amir.o dipeptide (293 mg, 0. 84 mmol~
To this was add~d Fmoc-~5et OPfp (450 mg, 0 . 84 mmol) :
f ollowed by the minimum ~uantity of ethyl ac~tate to gi~e a clear solutio~. The ~olve~t was immediately -:
removed and the resulting ~i~cou~ oil heated in a water bath at 65~C for 45 minut~s. Trituration of l:h~
2 0 resulting re3in with diethyl ether yielded the .
protect~3d tripeptide (418 mg, 78%).
Fmoc-Trp~Met Asp (OBut)--DL MePh~--N~2 The preGedins~ tripeptide ~300 mg, 0 . 43 mxnol) ~as N-deprotected u~in~ 20% piperidine in DME? ~or 20 minutes. ~emo~ral of the ~olvent followed by ~rituration o~ the residu~ with n-hexane le~t th~ free tripeptide (198 mg, 0 . 41 mmol) . To this was a ded ~moc-Trp~OPfp ~243 mg, 0 . 41 mmol) and sufficient ethyl acetate to gi-re a clear solution. The solvent wa~
immedia~ely remo~ed and the result~ ng mixture heated at 654C for 15 minutes. Trituration vf the residue with diethyl ether ga~re a white solid, 283 mg, 7896.
WO 92/192~3 PCr/l~S~2/030lS
210~7~3 Trp-Met-Asp-DL-MePhe NH2 CF3t::02H ( B ) .
The preceding tetrap~ptide was M-deprotected using 2û% piperidine in D~F for 20 minutes then the solvent ~:
remo~ed and the residue triturated with n-h~xane, lea~ring the fr~e amino tetrapeptide. This was treated ~-with thic~anisole and ethanedithiol (100-250 ,UL~mmol and 50-100 ~IL/mmol) followed by 95% aqueous TFA
(3-5 mL/mmol) for 5 minutes. All volatiles were remo~red and the residue triturated wikh diethyl ether, gi~ving a white solid, puri~ied by pr~parati~re ~ C.
Trp--~qet--DL--MeAsp--Phe-NH2 CP'3C02H (9) Fmoc-DL--MeAsp (OBut)--Phe--NH2 Fmoc--DL-MeAsp (0}3ut) 0Pfp ~1. 46 g, 2 . 47 mmol) and H-Phe-NH2 (O . 4~ g, 2 . 5 mmol) were dissolved in the :
mini~num quantity of ethyl acetate to gi~e a cl~ar solution~ then the ~301~rent ~s immediately res~oY~d.
The mixtur~ wa~ then heat~d at 100C for 1 hour and .-cool~d . Tritur~tion of the residue with diethyl ether ~ ~
yielded th protect~d dipepti.de, 993 mg, 71%. ~:.
Fmoc-~et-DI.-MeAsp (OBut)--Phe--NH2 .:.
The preceding dip~ptide (571 mg, 1. 0 mmol) was N-deprot~cted as pre~iou ly describad. Following trituration with n-hexane, the residue was mix~d with Fmoc-~et--OPfp (350 mgl 0 . 65 mmol) ar~d suffi ::ierlt ethyl ~ `:
acetate to dissol~re. Immediate removal of the ~ol~rent ~.
was followed by heatirlg the residue at 65C for 4 hours. Trituration of the residue with diethyl ether....~rielded the protected tripeptide, 210 mg, 4696.
A further crop could be isolated from the ethereal washinys by chromatography.
.... ... .. ..
WO 92/192~3 PCrfUS92/03015 2 1 ~ ~ 7 6~ ~34~ ` ~ ~
Fmoc-Trp-Met -Dl,-MeA~p ( OBut )--Ph-NH2 The above tripeptide (400 mg, 0.57 mmol) was N--deprotected as pre~riously described. Following trituration of the re~idue with n-hexane, 258 mg of ~-:
product were i~;olated. 200 mg (0.4? mmol) of this and Fmoc-Trp-OP:Ep (246 mg, 0 . 42 ~unol) ~as added ~ith ~ -sufficient ethyl acetate to dissol~e. The solverlt ~as immediately remo~ed and the residue heated at 65C fox .
2 0 minu es . Trituration of the resulting mixture with l 0 diethyl ether yielded the protected tetrapeptide, ~ -283 mg, 7796 . .
.
Trp-Metol~ Me~p-Ph~-NH2 CF3C02~ ( 9 ) The preceding tetrapeptide was treated as the tetrapeptide, yielding (8~ and gave after final trituration with ether.
.. ~
Claims (16)
1. A compound of formula I
or a pharmaceutically acceptable salt thereof wherein R1 is hydrogen, BOC, 1-Adoc, 2-Adoc, Z, FMOC, R12R13R14Si(CH2)mOCO- wherein R12, R13, and R14 are each independently lower alkyl of from 1 to 3 carbon atoms and m is an integer of from 0 to 3, R15CO- or R15NHCO- wherein R15 is a straight or branched alkyl of from 1 to 6 carbons or 1- or 2-adamantyl;
A is a bond, Gly, .beta.-ALA, GABA, DAVA, Aib, NH(CH2)p wherein p is an integer of from 1 to 6, NHCO, Met-Gly, Tyr-Met-Gly, or Asp-Tyr-Met-Gly, Asp-Tyr(OSO3H)-Met-Gly;
R2, R4, R6, and R8 are each independently hydrogen with the proviso that not all are simultaneously hydrogen, lower alkyl, -CH=CH2, C?CH, -CH2-CH=CH2, -CH2C?CH, -CH2Ar, -CH2OR, -CH2OAr, -(CH2)qCO2Rr, -(CH2)qN(R)2 wherein q is an integer of from 0 to 3, R is hydrogen or lower alkyl, Ar is a mono- or polycyclic unsubstituted or substituted carb- or heteroaromatic or hydroaromatic moiety;
R3 is 2- or 3-indolyl, 1- or 2-naphthyl, 2- or 3-benzothienyl, 2- or 3-benzofuranyl, 2- or 3-indazoyl or phenyl with from 0 to 3 substituents each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, methoxy, trifluoromethyl, nitro, hydroxy, NH2, and OCF3;
R5 is straight or branched lower alkyl of from 1 to 6 carbon atoms, (CH2)rS(CH2)sCH3 wherein r is an integer of from 1 to 6 and s is an integer of from 0 to 6;
n is an integer of from 1 to 3;
R7 is -OH, -COOH, tetrazole, triazole, -COOR16, -CONR10R11, wherein R16 is lower alkyl, and R10 and R11 are as described below;
R9 is selected from R3 above;
R17 is -?-NR10R11, CH2OR18 or H wherein R10 and R11 are each independently hydrogen, lower alkyl, alkoxycarbonyl, carboxyalkyl, or R10 and R11 togethrr form a ring of from 3 to 10 atoms with the nitrogen to which they are attached which ring contains atoms selected from carbon, nitrogen, oxygen, and sulfur;
R18 is R1 or CH2CO2H or CH2CONR10R11 or CONR10R11;
R2 and R3, R4 and R5, R6 and R7, R8 and R9 or R9 and R10 may together form a ring of from 3 to 8 atoms which atoms are selected from carbon, nitrogen, oxygen, and sulfur.
or a pharmaceutically acceptable salt thereof wherein R1 is hydrogen, BOC, 1-Adoc, 2-Adoc, Z, FMOC, R12R13R14Si(CH2)mOCO- wherein R12, R13, and R14 are each independently lower alkyl of from 1 to 3 carbon atoms and m is an integer of from 0 to 3, R15CO- or R15NHCO- wherein R15 is a straight or branched alkyl of from 1 to 6 carbons or 1- or 2-adamantyl;
A is a bond, Gly, .beta.-ALA, GABA, DAVA, Aib, NH(CH2)p wherein p is an integer of from 1 to 6, NHCO, Met-Gly, Tyr-Met-Gly, or Asp-Tyr-Met-Gly, Asp-Tyr(OSO3H)-Met-Gly;
R2, R4, R6, and R8 are each independently hydrogen with the proviso that not all are simultaneously hydrogen, lower alkyl, -CH=CH2, C?CH, -CH2-CH=CH2, -CH2C?CH, -CH2Ar, -CH2OR, -CH2OAr, -(CH2)qCO2Rr, -(CH2)qN(R)2 wherein q is an integer of from 0 to 3, R is hydrogen or lower alkyl, Ar is a mono- or polycyclic unsubstituted or substituted carb- or heteroaromatic or hydroaromatic moiety;
R3 is 2- or 3-indolyl, 1- or 2-naphthyl, 2- or 3-benzothienyl, 2- or 3-benzofuranyl, 2- or 3-indazoyl or phenyl with from 0 to 3 substituents each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, methoxy, trifluoromethyl, nitro, hydroxy, NH2, and OCF3;
R5 is straight or branched lower alkyl of from 1 to 6 carbon atoms, (CH2)rS(CH2)sCH3 wherein r is an integer of from 1 to 6 and s is an integer of from 0 to 6;
n is an integer of from 1 to 3;
R7 is -OH, -COOH, tetrazole, triazole, -COOR16, -CONR10R11, wherein R16 is lower alkyl, and R10 and R11 are as described below;
R9 is selected from R3 above;
R17 is -?-NR10R11, CH2OR18 or H wherein R10 and R11 are each independently hydrogen, lower alkyl, alkoxycarbonyl, carboxyalkyl, or R10 and R11 togethrr form a ring of from 3 to 10 atoms with the nitrogen to which they are attached which ring contains atoms selected from carbon, nitrogen, oxygen, and sulfur;
R18 is R1 or CH2CO2H or CH2CONR10R11 or CONR10R11;
R2 and R3, R4 and R5, R6 and R7, R8 and R9 or R9 and R10 may together form a ring of from 3 to 8 atoms which atoms are selected from carbon, nitrogen, oxygen, and sulfur.
2. A compound according to Claim 1 wherein R1 is Fmoc, Z, Boc, or Me3Si(CH2)2OCO2H;
A is a bond, Gly, .beta.-ALA, Asp-Tyr-Met-Gly;
R2, R4, R6, and R8 are each independently alkyl of from 1 to 3 carbon atoms, carboxyalkyl of 1 or 2 carbon atoms, or CH2OH;
R3 is 3-indazolyl, 3-benzothienyl, 2-benzofuranyl, 3-benzofuranyl, or 2-bromo-3-benzofuranyl;
R5 is isobutyl, sec-butylethyl, n-propyl, or i-propyl;
R7 is CONH2, CONMe2, or OH;
R9 is selected from R3;
R17 is -?-NR10R11, wherein R10 and R11 are each independently alkyl of from 1 to 3 carbon atoms or together R10 and R11 form a ring of from 4 to 8 atoms including the nitrogen to which they are attached.
A is a bond, Gly, .beta.-ALA, Asp-Tyr-Met-Gly;
R2, R4, R6, and R8 are each independently alkyl of from 1 to 3 carbon atoms, carboxyalkyl of 1 or 2 carbon atoms, or CH2OH;
R3 is 3-indazolyl, 3-benzothienyl, 2-benzofuranyl, 3-benzofuranyl, or 2-bromo-3-benzofuranyl;
R5 is isobutyl, sec-butylethyl, n-propyl, or i-propyl;
R7 is CONH2, CONMe2, or OH;
R9 is selected from R3;
R17 is -?-NR10R11, wherein R10 and R11 are each independently alkyl of from 1 to 3 carbon atoms or together R10 and R11 form a ring of from 4 to 8 atoms including the nitrogen to which they are attached.
3. A compound according to Claim 1 wherein R1 is hydrogen or BOC;
A is a bond or .beta.-ALA;
R2, R4, R6, and R8 are each independently CH3 or CH2CO2H;
R3 is 3-indolyl, 1-naphthyl, or 2-naphthyl;
R5 is n-butyl, n-pentyl, (CH2)2SCH3 or (CH2)2SCH2CH3;
R7 is OH, tetrazole, triazole, or CO2H;
R9 is phenyl, 1-naphthyl, or 2-naphthyl, R17 is -?-NR10R11, wherein R10 and R11 are each independently hydrogen, CH3 or together R10 and R11 form a ring of 5 or 6 atoms including the nitrogen to which they are attached.
A is a bond or .beta.-ALA;
R2, R4, R6, and R8 are each independently CH3 or CH2CO2H;
R3 is 3-indolyl, 1-naphthyl, or 2-naphthyl;
R5 is n-butyl, n-pentyl, (CH2)2SCH3 or (CH2)2SCH2CH3;
R7 is OH, tetrazole, triazole, or CO2H;
R9 is phenyl, 1-naphthyl, or 2-naphthyl, R17 is -?-NR10R11, wherein R10 and R11 are each independently hydrogen, CH3 or together R10 and R11 form a ring of 5 or 6 atoms including the nitrogen to which they are attached.
4. A compound named .alpha.-methyl-DL-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
5. A compound named .alpha.-methyl-D-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
6. A compound named .alpha.-methl-L-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
7. A compound named N-[[(1,1 dimethylethoxy)-carbonyl]-.alpha.-methyl]-L-trytophyl]-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
8. A compound named N-[(1,1-dimethylethoxy)-carbonyl]-.alpha.-methyl-DL-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-.alpha.-phenylalaninamide.
9. A compound named L-tryptophyl-2-methyl-DL-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
10. A compound named N-[(1,1-dimethylethoxy)-carbonyl]-.alpha.-methyl-D-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
11. A compound named glycyl-.alpha.-methyl-D-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninasnide.
12. A compound named glycyl-.alpha.-methyl-DL-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
13. A compound named glycyl-.alpha.-methyl-L-tryptophyl-L-methionyl-L-.alpha.-aspartyl-L-phenylalaninamide.
14. A phanmaceutical composition comprising an amount of a compound aecording to Claim 1, effective to suppress the appetite in a mammal, and a pharmaceutically acceptab1e carrier.
15. A method of suppressing appetite in a mammal, comprising administering an effective appetite suppressing amount of a compound according to Claim 1.
16. A compound selected from:
Fmoc-MePhe-NH2, Fmoc-Asp(OBut)-MePhe-NH2, Fmoc-Met-Asp(OBut)-MePhe-NH2, Fmoc-Trp-Met-Asp(OBut)-MePhe-NH2, Asp(OBut)-MePhe-NH2, Asp-MePhe-NH2, Fmoc-Met-Asp-MePhe-NH2, Met-Asp(OBut)-MePhe-NH2, Met-Asp-MePhe-NH2, Trp-Met-Asp-(OBut)-MePhe-NH2, Fmoc-Trp-Met-Asp-MePhe-NH2, Fmoc-MeAsp-(OBut)-Phe-NH2, Fmoc-MeAsp-Phe-NH2, Fmoc-Met-MeAsp(OBut)Phe-NH2, Fmoc-Trp-Met-MeAsp(OBut)-Phe-NH2, MeAsp(OBut)-Phe-NH2, MeAsp-Phe-NH2, Met-MeAsp(OBut)-Phe-NH2, Met-MeAsp-Phe-NH2, Fmoc-Met-MeAsp-Phe-NH2, Fmoc-Trp-Met-MeAsp-Phe-NH2, MeMet-Asp(OBut)-Phe-NH2, Fmoc-MeMet-Asp(OBut)-Phe-NH2, Fmoc-MeMet-Asp-Phe-NH2 MeMet-Asp-Phe-NH2, Fmoc-Trp-MeMet-Asp(OBut)-Phe-NH2, Fmoc-Trp-MeMet-Asp-Phe-NH2, Fmoc-MeTrp-Met-Asp(OBut)-Phe-NH2, Fmoc-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-Trp-Met-Asp-MePhe-NH2, Fmoc-Gly-Trp-Met-Asp(OBut)MePhe-NH2, Gly-Trp-Met-Asp(OBut)MePhe-NH2, Fmoc-Gly-Trp-Met-MeAsp-Phe-NH2, Fmoc-Gly-Trp-Met-MeAsp(OBut)-Phe-NH2, Gly-Trp-Met-MeAsp(OBut)-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Fmoc-MeMet-OH, Fmoc-MeMet-OPFP, MeAsp(OBut)OH, Fmoc-MeAsp(OBut)OH, and Fmoc-MeAsp(OBut)OPFP.
Fmoc-MePhe-NH2, Fmoc-Asp(OBut)-MePhe-NH2, Fmoc-Met-Asp(OBut)-MePhe-NH2, Fmoc-Trp-Met-Asp(OBut)-MePhe-NH2, Asp(OBut)-MePhe-NH2, Asp-MePhe-NH2, Fmoc-Met-Asp-MePhe-NH2, Met-Asp(OBut)-MePhe-NH2, Met-Asp-MePhe-NH2, Trp-Met-Asp-(OBut)-MePhe-NH2, Fmoc-Trp-Met-Asp-MePhe-NH2, Fmoc-MeAsp-(OBut)-Phe-NH2, Fmoc-MeAsp-Phe-NH2, Fmoc-Met-MeAsp(OBut)Phe-NH2, Fmoc-Trp-Met-MeAsp(OBut)-Phe-NH2, MeAsp(OBut)-Phe-NH2, MeAsp-Phe-NH2, Met-MeAsp(OBut)-Phe-NH2, Met-MeAsp-Phe-NH2, Fmoc-Met-MeAsp-Phe-NH2, Fmoc-Trp-Met-MeAsp-Phe-NH2, MeMet-Asp(OBut)-Phe-NH2, Fmoc-MeMet-Asp(OBut)-Phe-NH2, Fmoc-MeMet-Asp-Phe-NH2 MeMet-Asp-Phe-NH2, Fmoc-Trp-MeMet-Asp(OBut)-Phe-NH2, Fmoc-Trp-MeMet-Asp-Phe-NH2, Fmoc-MeTrp-Met-Asp(OBut)-Phe-NH2, Fmoc-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-Trp-Met-Asp-MePhe-NH2, Fmoc-Gly-Trp-Met-Asp(OBut)MePhe-NH2, Gly-Trp-Met-Asp(OBut)MePhe-NH2, Fmoc-Gly-Trp-Met-MeAsp-Phe-NH2, Fmoc-Gly-Trp-Met-MeAsp(OBut)-Phe-NH2, Gly-Trp-Met-MeAsp(OBut)-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp-Phe-NH2, Fmoc-Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Gly-MeTrp-Met-Asp(OBut)-Phe-NH2, Fmoc-MeMet-OH, Fmoc-MeMet-OPFP, MeAsp(OBut)OH, Fmoc-MeAsp(OBut)OH, and Fmoc-MeAsp(OBut)OPFP.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US69075491A | 1991-04-24 | 1991-04-24 | |
| US690,754 | 1991-04-24 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2106765A1 true CA2106765A1 (en) | 1992-11-12 |
Family
ID=24773822
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002106765A Abandoned CA2106765A1 (en) | 1991-04-24 | 1992-04-15 | Cck analogs containing ó-substituted amino acids |
Country Status (8)
| Country | Link |
|---|---|
| JP (1) | JPH06506941A (en) |
| AU (1) | AU1879892A (en) |
| CA (1) | CA2106765A1 (en) |
| IE (1) | IE921320A1 (en) |
| MX (1) | MX9201883A (en) |
| PT (1) | PT100427A (en) |
| WO (1) | WO1992019253A1 (en) |
| ZA (1) | ZA922951B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5814316A (en) * | 1994-08-05 | 1998-09-29 | Wisconsin Alumni Research Foundation | Compound to mimick a naturally occurring peptide's effect |
| GB0105069D0 (en) * | 2001-03-01 | 2001-04-18 | Univ Ulster The | Modified peptide |
| US20040052928A1 (en) | 2002-09-06 | 2004-03-18 | Ehud Gazit | Peptides and methods using same for diagnosing and treating amyloid-associated diseases |
| US7491699B2 (en) | 2002-12-09 | 2009-02-17 | Ramot At Tel Aviv University Ltd. | Peptide nanostructures and methods of generating and using the same |
| WO2004060791A1 (en) | 2003-01-07 | 2004-07-22 | Ramot At Tel Aviv University Ltd. | Peptide nanostructures encapsulating a foreign material and method of manufacturing same |
| JP5137400B2 (en) | 2003-06-30 | 2013-02-06 | テル アヴィヴ ユニヴァーシティ フューチャー テクノロジー ディヴェロップメント エル．ピー． | Peptides for diagnosing and treating amyloid-related diseases, antibodies thereto, and methods of use thereof |
| JP4917889B2 (en) | 2003-09-25 | 2012-04-18 | テル アヴィヴ ユニヴァーシティ フューチャー テクノロジー ディヴェロップメント エル．ピー． | Compositions for treating amyloid-related diseases and methods of use thereof |
| WO2006018850A2 (en) | 2004-08-19 | 2006-02-23 | Tel Aviv University Future Technology Development L.P. | Compositions for treating amyloid associated diseases |
| WO2007043048A2 (en) | 2005-10-11 | 2007-04-19 | Ramot At Tel Aviv University Ltd. | Self-assembled fmoc-ff hydrogels |
| BR112013012033A2 (en) | 2010-11-15 | 2016-08-09 | Merz Pharma Gmbh & Co Kgaa | dipeptide analogues for the treatment of conditions associated with the formation of amyloid fibril |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NZ218607A (en) * | 1985-12-19 | 1989-10-27 | Pennwalt Corp | Tri-to acta-peptides with sulphate ester groups: obesity treatment |
| US5013722A (en) * | 1986-06-06 | 1991-05-07 | Hoffmann-Laroche Inc. | Cholecystokinin analogs for controlling appetite |
| US5098995A (en) * | 1987-05-22 | 1992-03-24 | The Salk Institute For Biological Studies | GRF Analogs VIIA |
-
1992
- 1992-04-15 JP JP4510283A patent/JPH06506941A/en active Pending
- 1992-04-15 WO PCT/US1992/003015 patent/WO1992019253A1/en active Application Filing
- 1992-04-15 AU AU18798/92A patent/AU1879892A/en not_active Abandoned
- 1992-04-15 CA CA002106765A patent/CA2106765A1/en not_active Abandoned
- 1992-04-23 ZA ZA922951A patent/ZA922951B/en unknown
- 1992-04-23 IE IE132092A patent/IE921320A1/en not_active Application Discontinuation
- 1992-04-23 MX MX9201883A patent/MX9201883A/en unknown
- 1992-04-24 PT PT100427A patent/PT100427A/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| AU1879892A (en) | 1992-12-21 |
| PT100427A (en) | 1993-08-31 |
| JPH06506941A (en) | 1994-08-04 |
| IE921320A1 (en) | 1992-11-04 |
| MX9201883A (en) | 1992-10-01 |
| ZA922951B (en) | 1993-10-25 |
| WO1992019253A1 (en) | 1992-11-12 |
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