CA2275442A1 - Melanocortin derivatives for specific binding of melanocortin receptor 3, 4 or 5 - Google Patents
Melanocortin derivatives for specific binding of melanocortin receptor 3, 4 or 5 Download PDFInfo
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- CA2275442A1 CA2275442A1 CA002275442A CA2275442A CA2275442A1 CA 2275442 A1 CA2275442 A1 CA 2275442A1 CA 002275442 A CA002275442 A CA 002275442A CA 2275442 A CA2275442 A CA 2275442A CA 2275442 A1 CA2275442 A1 CA 2275442A1
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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
- C07K14/665—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
- C07K14/68—Melanocyte-stimulating hormone [MSH]
- C07K14/685—Alpha-melanotropin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention provides novel peptides derived from the group of hormones including ACTH. These so-called melanocortins can target different receptors which often have different localizations on several tissues. The presently invented peptides are useful for targeting receptors in the nervous system in an agonistic manner. Pharmaceutical compositions and uses are also disclosed.
Description
MELANOCORTINS
The present invention relates to t:he field of melanocortin peptides.
Melanocortins (which used to be called melanotropines also) are peptides originally derived from a larger precursor protein named pro-opiomelanocortin. The natural melanocortins share the heptapeptide core sequence Met-Glu-His-Phe-Arg-Trp-Gly. These melanocortins include cK-MSH (cx-melanocyte stimulating hormone), (3-MSH, y-MSH, y-LPH (y-lipotropin hormone) and ACTH (adrenocorticotrope hormone).
Melanocortins have a wide range of: biological activities. They have been known for a long time to stimulate pigmentation and corticosteroidgenesis, but they hive also been shown to induce excessive grooming behaviour in the rat, to stimulate conditioned active avoidance response, to increase blood pressure and heart rate, to accelerate nerve regeneration and to modulate immune responses. Quite recently five neuropeptide receptors for melanocortins have r>een identified and cloned.
These receptors have different di~;tribution patterns (in presence as well as in abundance) over the different tissue types. They belong to the family of so-called G-protein coupled receptors. Melanocortin rs~ceptor 1 (MCR-1) is expressed in melanocytes, whereas MCR-2 is the ACTH receptor expressed in for instance the adrs~nal gland. Melanocortin receptors 3, 4 and 5 have been found to be expressed in the central nervous system. The cognate ligands of these receptors have profound neuropharmalogical effects of, such as facilitated arousal, motivation, attention, memory and learning. The ligands have also been implicated in food-motivated behaviour. Further a relation with antipyretic activity has been disclosed.
Many different (synthetic) analogs of melanocortins have been . prepared and suggested to be of use in activating or blocking one or more of the MC-receptors. Z~his agonistic or antagonistic action has then been suggested to be useful in applications relating to pigmentation, nerve regeneration, etc.
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However, the melanocortins developed so far lack n .
specificity (selectivity) for the receptor* expr0 sed in the nsxvous system, and/or they lack ,i.n suffici~nt b ding .~ affinity or capability to,induce the receptor med at~d respoase or to block said response. Typically it ld ba desired. that a drug targeting the MC-raoeptors ba highly potent, orally adma.nistexabla, r~~asonalily resista~t to breakdaarn in the body (have a ~uf~Eicient half-lif~) cad able to cross the blood-brain barrier.
..
l0 The present invention provides peptides or peptid -like structures that meat the structural requirements o be useful as essentially 1tC-receptor specif;i.c drugs.
Thus th~ invention provides a pap~tida having spsc~.fic binding affinity for a melanocortino rec~ptor, preferabiylthe mc3, mc4 or mc5 recaptox conprisiag th~ sego~nve ~ .
X-Y--His-(D-2-Thienyl-Ale)-ar9-(Z) or x-x-His- ( 3-.pyridyi-D-Al~a ) -Arq- ( ~ ) whereby X and Y a~ amino acid residues and $ is Ian aromatic amino acid residue. , According to the prest~nt inv~entioxi the above po ides era aqonists for MsH aotivity ~rhfch as.~ highly potan. A vest' 25~ isportant contributioa to the high potency attributsd can to the 7-position (oountvd gut in t:h,~ oriqinsl -moiaculsr,) A
which should bs D-2--thianyl~Ala o~~ 3-pyridyi-D- a and for which only wty limited and very similar residue may be substituted without losi~xg the inc~raaso in aqonit potency.
AnothQr important contribut3.on to M88 agonistict3.Wity i*
the omission of residu~s 1-3 and/a~r the o~n3ssiono~ rasidusa 11-13. A hiqh.contribution to activity is also ovidad by p the pretence of an aromtic amino acid. residua at po ition~9.
positions 4 and 5 should normally not bs omitted these residuts should ba pras~nt though it is tar critical lass wdich aaa~no acid residues era preson~t at said itions. rt is clear that at least consarrvad substitutions era ilow*d for these positions, but also lass conserwd substitu tions grill 9. IorJ, 1999 s ~ ; 9 ~ ~~~~ ocfipo~~ ~ ~o~E~~Ty (3 i ) 3 ~422?39 ~ No. 4345 p. 19 3 ~ . .
histidine residue at position 6 .and the arginine esidve at position 8 ar~ quite important for activity and ~ ould only be replaced by very consorvatiw suJbstitutions, if a ~a11.
5~~ Especially the more important residues in general should not be. all replaced by substitutes i~z on~ and the sam~ molecule.
The preferred residue at position is Naftyi-alaai#ie, bo it. in the D-or is the L-configuration.
The presenc~ of this residue 1~ads to a fuxth~r i~ncreae~e i.r~
potency.
An a~n.W o acid residue at position 10 is not esse~ial. for the activity of the molecul~, but it does seem to have some effect. If a residue fs present j.t is pref~rred t,~at this residue is Qlycine or lysine, whe~rlby the latterihas the additional. advantage that it provides a reactive ;moiety whicb can be used to couple the peptids~ tv other molecules or to make the peptide cyclic. In the e~wnt that a cyc is paptida is to be produoed, which~is preferzed, since the ha f~lit~ of such a cyclic peptide is improved over the half-~ifo of the lin~ar fo~n, then a reactiv~s moiety at the other~end shouxd also be provided. Another advantage of having a ~yalic paptida is that these peptides tend to have a higher sal~axivity for MC-receptors, in particular an disulphide bridge'inareasee the selectivity for the MC-4 receptor. Cyclic peptid~s oan ho~nsyer also be produced by providing rea~"tivs moieties tside the essential sore that enable closury! of the loo ~uch ax p~ ~ ' reactive moieties leading to a lacaam.
The preferred residues it positions X and x, (nr~~ninq 4 and 5 in the original ~CTB-core) are mle for x and G1y! or lisp for 7r, whereby the presence of Asp loads to a further alivantage in having a raaatiw moiety for making a lactam.
The peptides.accordinq to the invention are gen~aliy more potent than irt~a itself. The prefQx~red peptides h~w poteneias of up to 100 timQS the potency of leg. Less pots t are within the scope of the present inwntioa, s ncelpoteency' ~ is not the only criterion which is required for ~a successful peptide-based drug. As already mentioned, half-l~~ife arid selectivity are also important par,~ters.
g~nerally not lead to'a significant loss of poten~y. The 9, dU~d. 1999 15; Ol ~'ER OCTR~JOIBUREAUX (3i ~ 334227319 . N0, 4345 P. 20 4 ~ , , . , . The tests used for~detes~aining the potency of thelpeptides are .
the regular in itro t~ate as well as an i vivo ~est i,n rats whereby the growing behaviour i;s measured: The.r~eults in a -groos~ng assay are good indications that the.pept~.des will be 5' able to activate the receptor and thereby the G-protein cascad~ coupled to said receptor and thus the pepltides can be used as agonists for ~sC-receptors . Targeting the ~.-receptor in particular of the MC..4 rec~pt«r fox which a t~cularly selective peptide has been provided by the pres invention in an agonistic manner is useful in the treatment of Ci~s-disorders, neurophat6ies, obesity, and in particuilar for diabetes related neuropathy, as 'sell as neuropat~y ~ a result of cytotoaic treatments (chemotherapy aird the if.ke ) .
Th~retore tho pres~nt invention also provides ph~rmscantical co~ositic~ns capable of treating the above conditions. Dosages for suoh treatments will usually be given once n~day in doses of about 1 erg to about 100 mg p~z~ dosage unit, preferably 10 erg - 10 mq, more preferably 5b~ uq - 1 mg. The~dosage should ZO zmsult is a concentration in the body of between!0,1 nay at~d 1 um. preferably 1 n~1 - 100 n~i, aaost preferably ~0 - 50 nl~l.
The compositions msy comprise the usual additivs~ for usual dosage formats for peptide drugs or for peptide ~leriv~sd drugs, The format is preferably an oral formulation :ua~ as s tablet, Z5 a granulate, a powder or a~liquid formulation, a~though enteral and parerrteral administrations may find ~ppiication as wail. particularly preferred are ~~o~positions~ wherein a peptide according to the xrrventio~a is combined w~.th a drug aiuiing to prevent or that leads t~~ neuropathy, s~ch as~insulin 3 0 arrd cytotoxio agents . ' The invention w~.ll be explained in spore detail the following eape~r3,m~enta3, part .
I
i 9. JU?u', 1999 13. ~2 VER OCTRO~aISLTREAUK (31; 3342273:9 NG, 4343 P, 2I
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Material sad ~tstbods ' l~le,Ianocortin Ifgand receptor activity 5 v guman embryonal kidney (8810 293) cells mere stabi~r transfected with the human MC3(Gantz et al. ~tBC 1993. 268a824~6-8250), human MC4 (Gaeta et al. 1993. JHC; 268a15174015179~) or human MC5 receptor constructs using the calcium ph4spha~t precipitation method. A6 a reporter plasmid 10 ~ ~ of the pCRE-LaGz vector (Chon tt al. 1995. Anal.Biochem. ZZ6~349-354), in 'vrhich a c~ responsiv! element drives ezprsssio~ of the Lacx gene, was transfected at ttu begj.aning of each ~~rioent. The day after pCRE~LacB transfection, cells were split in 96-wells plates.. After 48 hours cells were stimulated with var5ring concentrations of the MC recepto=~ ligands in ass~y miodium Dl~t t 0 :1 mg/ml HBA, 0 . i mi~~ ZBMx ) for 6 house . ' Cells were lysed in lysir buffer_by a freQae-thaw round, suk~stratQ buffer (60~! sodium phosphate, i mt~t MqClZ) 5 m!t ø-merea~tosthanol, 200 NQ/ml ONPG) was added and col.ls were incubat~rd at 37~C for 1 hour. The activation of the cAll~ signal~transd~ction pathway upon receptor activation was detected in a micro;plat~ reader (8forad le'del 3500) a 405nm using a colorimetri~ assay as dQecribed by Chen et a1.) Animals, 3aqpiantatson o! carrnctlar~, fntracerebrov$ritricti3ar injection .
dale ~Pistar rats woigt~ing 120-130 g were implant~d with cannulas into the foramen intrave~ntriculars uade~r hypno~o anaesthesia (8rakkee at al., Life 3cieace'vol. ih 19'79, ).
hats were allomsd to recover for .3 days and usodj for experiments during the neat 10 da;rs. In case that rats were used for more than one grooming e;~perimrat they ere allomsd to recover for at least 3 days between subsequo~itwxpe~riments, peptides (15 nq)~dissolvod in 3 N~~ saline (154 m~ NaCl) were injected i.c.v. by means of a Ha~~lton syring~s. 't3ruom,ing taste ' were performed according to (Gispc~n~ et al, Lab. ~llnim. Sci. 29 1975). Rats were placed into the obsorvatioa boos ima~ediately after the infection. obsezvaticn started 15 miniafter the i i CA 02275442 1999-06-16 j I
u. JUN, !9~'~ _~ 5: Q? UER UC'fROnBUREAUX i3!' 3~~4~~'l3' ~ 1VU, 4~?4'~
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injection and continued for 50 min. Grooming was cored each 15 sec over 50 min, thus the msxa.mal grooming scope for a rat is 200. ~ j _ 5~ syntbl,ia of peptides Pur'~f i ca -i on o ~~ ~ ' Preparative BpLC was carried out using a eaters P~sp 4000 .
liquid chromato9raph, eguipped wj.th a Waters aCM ~modui~ with two prepBak cartridges plus c~uardl cartridge ( 25:2~.0 mm) filled with Delta-Pak C18 material. Peptides tm~re detec~ad at 230 nm using a Waters 486 spectrophoton~a~ter with a prap~rativrr cell.
purifications wexe performrad in g~radfonts a*in9 v~ater with 0.11 trifiuoroacetic acid (TFA) and aaetonitrile.with O.lt TPA.
.1S i l~tettiods for synthesis and cyclisation of a-r~iSH p~ptides a ~ltinl~es~.id~ Svnthnw~t*
We .used a eanilton 1r11crolab 2200 to synthesise u~ to 40 peptides sinaltaneously at 30 ~1 scan. Thi 8a~iiton Microlab 2200 xaa programesd to delivor washing ~olveats and roagents to a rack with 40 individual 4 ml columsi* with filter, containing Rf.dk ( 4- ( 2 ~ , 4,' .-dinsthosypheny~-I s'nrx-aminos~hyl)-pheno:y) resin for peptide *ynthssf.~. The columns wets drained automatically after ~eaoh atop by vacuum. The 2S coupling Gyclo.rras based on Fmoc/»BTV (2-(1H-ben~otria~sol-1-yi)-1,1,3.,3-tetraa~thyZuronium hoaafluorophospha~e) chemistry [Fields ~t al.~ Peptide Research ~~, 95-101] usin~ double coupling steps of 40 minutes. Pep~tidos ire depr~tec~ted and cl~avod in 2 hre using 1.5 ml of as mixture o! tr~fiuoroacvtic acid/phenol/thioanisolelwater/ethanedithioi j 10/0.7510.5/0.5/0.25 and then precipitated twicelby adding hr~ane/disthy3.ether 111. The precipitate was dri~d and lyophi7.ised from water/acetoaitri7:e. , C~~~,n_ of n nt; ~. .
PLC purified' or crnd~e peptides wore used for cyriization. via a disulfide bridge with cysteiaes or via a lactajs br3,.dQo w3.th the side ehaias of aspartio acid and lysiaes~ !
A. disulfide bridge: crude MBJ06 (40 mg) was dissolved in 40 ml of a 0.5~ ammoniumbicarbonate~ solution at pH 8 and stirred overnight. After 24 hours no free sulphydryl groups were detected using Ellman's reagent and the product was lyophilized after addition of 0.5 ml of acetic acid. The y peptide was dissolved in 3 ml of 40~ acetic acid and purified by preparative HPLC in a gradient of 14$ to 21$ acetonitrile in water (containing 0.1~ TFA) i.n 30 min. Yield after purification: 22.8 mg.
B. Lactam bridge: a mixture of 20 ml of DMF (peptide grade), 26 mg of Benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP, 0.05 mmol) and 0.017 ml of DIEA
(0.1 mmol) was added to crude MH~J07 (20 mg, 0.012 mmol). The cyclization reaction was followed by analytical HPLC. After 2 . hours the mixture was acidified to pH 4 using 0.1 M HC1. The product was purified by preparative HPLC after dilution with 30 ml of water in a gradient of 21~ to 30~ acetonitrile in water, containing 0.1~ TFA, in 30 min. Yield 15.4 mg.
Results Screening of modified MSH peptides generated by PEPSCAN~
revealed several amino acids that increased MSH potency. D-2-thienyl-Ala and 3-pyridyl-D-Ala at position 7 of the MSH
peptide was the most potent contributors of increased MSH
potency. Naftyl-Ala at position 9 also increased MSH potency (figure 1b). Deletion of positions 1-3 and 11-13 further increased MSH activity. Figure 1 shows the dose-response curves for the following peptides:
MBU 23 *2GH6R7G# linear MBU 24 *2GH6R8G# linear MBU 27 *CGH6R8C# cyclic disulphate MBU 28 *2DH6R8K# cyclic lactam MBU 29 *2DH6R7K cyclic lactam * acetyl # carboxamid 2= Norleucine, 6= D-Thienyl, 7= 2-naftyl-L-alanine, 8= 2-naftyl-D-alanine 9. 1999 1;03 VER OCTROOIBUREAUX ('j1) JUN.3~4~~'~31~
' Nu, 434 r .
. 8 ( .
.
. , , (figures Positions 4~and 5 may not be critical 3 sand 4) -At position 9 an aromatic acid (1~,H,Y)is highly ~reterred figure 2 ) . Figures 3 to 9 show ithe ~ffect' of the' different -amina acid substitutions'.at positions4 to 10 of ale-I~cSS, 5' xespeatively.
. I
. induction The activity of these peptides can ref 9roodi.ng the behaviour ~ following intracer~hro~rentricular $tions is.
in jec shown in figure 6. 1500 nQ is thr~
lowest dose of iMBH that induces excessive grooming ~behav~Lour.The five cQmpouads tested hers are 100x pore potent than Ms8 in viva.
Table 1. Calculated 8C50 values f:or the three di~ferQnt cell lures . ' hHC3 hMC:4 hl~lC~
l~tJ 23 5.510-9 ~.~i,~0-9 a.dr i . 4.80-$
i~u 24 3.010-9 1.?v10-8 , i I~1 Z7 I.3I0-~ 6.G~10-g 2~.5a a-s MeU 28 6.310-7 1.710 8 4.5~0~8 i ' I~1 24 . 1.410-9 ~ 1.510-8 2.3~,Q~8 i a-MSH 1.410-a 2.610-8 .3.0x0'"$
i Legends ' 3o Pi~urs 1. Dose response curves ~:or the differsr~t ldSB peptides on eBlC 293 cells aabiy e=pressing husr~n MC3, KC~4 sad MC5 receptors. M8U 27 shows specificity for the HC4 ;receptor. 1~U
23 is a potent li9aad on all three receptor .
~ignre 1b. effect of the different aainoacids ~.t position 7 and 9 togeth~e= with MHtI 23 and IKB~J 24. All peptides wero tested at 100 ruK oa a8x 293 cells stabiy expreseinq the three different 1~C receptorsf MC3, MC4 ~~~d MC5 rtoieptc~rs. values i I
9. JUA, 19u ~ '~ ~, ; f?? E'R'R OCTR~JG i BURE~~n (3', i 334??"3~ ~ N0. 434 P, a ' ~ 9 '' ' i represent the percenteg~ activit~~ as cao~pared to the maximal acti~rity of 100 n1~1 8n1?-MBH.
' Eiguro ~ . 8ffect of sini~le aminoacid Bubstitutiqns at 5~ aminoacid position number.9 of t~P-MS8. On the x-p~cis the aminoacids replacing~the endogenous aminoacid (Try) are depicted. values represent~th~a percentage activity as cospared to thQ maximal activity of 10 nM at-MISH. All pept'~des were tested at 10 nlK on H81C 293 cells stably ~apress~g the. three different 1KC receptorsfi~C3,~ MC4 s.nd L~t05 receptors. at this position on an NDp-M8g background. an aromatic amino acid ie higtily preferred.
i~iqure 3. affect of single ami,noacid substitnti~tss at aminoacid position numbtr 4 of Nle-MSg. On the Xrtazis the aminoacid~t replacing the endogenous aminoacid (lit) err depicted. Values represent the pert~ntag~ activity as comparod to' the masimal activity , of 100 nl~!', a-M88. A11 peptides were tested at 100 aM oa 8810 293 cells etably eapress~ng the. three different NC receptors= i~C3, MC4 and' 1~C,5 receptors.
FiQnre ~. 8ffect of single aminoacid substitutilons at aaiinoacid position numtior 5 of l~Ie~1i88. On tho x~-asis the aminoacids replacing the endogenous aa~inoaaid (G~.u) are depicted. Values represent the percentage activity as cvosparetl to ttLe ma~cfaval activity of 100 nl! a-1~3H. All pep~ti.des were .
tasted at 100 n1~ on HBx 293 cells stably a:pressiing t1~ three diffQrent 1KC recaptorsi MC3, KC4 and MC5 rec~pto~rs.
Figure 5. 8tfect of singlo aminoacid substitutions at aminoaeid position number d of Nl~e-MBH. 0:1 the 7~-eels the as~inoacids replacing the endogenous aminoacid (bis) are depicted. valves represent the perc~ntagQ activ~:ty as compared to the ma:imsl activity of 100 nM a--MS8. rrll peptides mere tested at 100 nip op B8R 293 calls stably a=pressing the three different 1~C reoeptors=MC3, MC4 and- ICS receptors.
i q. JUIV, 1996 i~~:~3 VER ~.CTR;'~CrBURE"L''r 31; 3 ?""
r s 3~~ c r 319 ~ NC-, 435 F. 26 . 10 . ' Fignre '6. Effect of ~ single aminoacid substitutions at aminoacid poeitiou number 7 of N7.e-MSH. On the X-!axis the aminoacids replacing the endogenous aminoacid (Pte) are depicted. Values represent the perc~atage activity as coa~ared to th~ maximai'activity of 100 nr!t a-l~H. A11 pep#id~s were tested at l00 nli on eSE 293 call~i stably ezpress~lag the three different MC receptor:; MC3) MC4 and HCS racapto~s. ~ -Pfgure ~. Effect of single aminoacid substitution: at amfnoacid position n~ober 8 of Nl,e-1188. Ori the Z+axis the ~ami.noacids replacing the endogenous aminoacid (A~g) airs depiotrd. values represent tho percentage activity as comparod to the maximal activity of 100 n~!t a-M88. 7111 peptides arere tosted ~ at 100 ate on B~ 293 cell~~ stably expressing tlu~ three different MC receptors;MC3, MC4 and 1~C~ receptors. Only fiv~
aminoacid~substitutioria mere reefed.
Figure o. Hffect of single amincxcid substitutibris at aainoacid position nunber 9 of Nl.e~MSB. oa tho 3~aais the aminoacids replacing tho tr~dog~nous ~0.i.noacid (Tlrp) are dQpiotod. Valuos~repreetat the pa~rcantage activity as compared to the ma~zimal activity of 100 nl~t~ a-M88. 1111 ptpitides were tested at 100 nH on 881 293 cells aably eupross~iaQ tho tdrQe d~,ftorerit iIC-rlceptorap M~C3, KC4 and MC5 receptors.
, -Figure !, affect of single amincxoid substitutions act aoinoacid position numba,t 10 of 1~'1Q-MBH. 0n the~X-a:is t~
aminoacids replaci~aq the endogenous anninoacid ( (~ly ) are depictod. values represent the percentage activity as co~mparod to thc~ mazimel activity of 100 nil a-~tSH. 1111 ps~tides were ttst~d at 100 ~u~t ~ on 293 collr stably e:pres~ting the three different MC' receptors ~ ~IC3, Mc4 and xC5 receptors .
rigur~ lo. .
Peptide ~faductd groaioiag ~ .
' 3 ;ri saline, ~or 3 ;rl saline with ~eithor 15 ng 1~Q peptides or 190 ng a-l~lBH or 1500 ng a-~18s wau in jeoted i.c.v. in rats and grooming behaviour was snored (mean t s.d.).
The present invention relates to t:he field of melanocortin peptides.
Melanocortins (which used to be called melanotropines also) are peptides originally derived from a larger precursor protein named pro-opiomelanocortin. The natural melanocortins share the heptapeptide core sequence Met-Glu-His-Phe-Arg-Trp-Gly. These melanocortins include cK-MSH (cx-melanocyte stimulating hormone), (3-MSH, y-MSH, y-LPH (y-lipotropin hormone) and ACTH (adrenocorticotrope hormone).
Melanocortins have a wide range of: biological activities. They have been known for a long time to stimulate pigmentation and corticosteroidgenesis, but they hive also been shown to induce excessive grooming behaviour in the rat, to stimulate conditioned active avoidance response, to increase blood pressure and heart rate, to accelerate nerve regeneration and to modulate immune responses. Quite recently five neuropeptide receptors for melanocortins have r>een identified and cloned.
These receptors have different di~;tribution patterns (in presence as well as in abundance) over the different tissue types. They belong to the family of so-called G-protein coupled receptors. Melanocortin rs~ceptor 1 (MCR-1) is expressed in melanocytes, whereas MCR-2 is the ACTH receptor expressed in for instance the adrs~nal gland. Melanocortin receptors 3, 4 and 5 have been found to be expressed in the central nervous system. The cognate ligands of these receptors have profound neuropharmalogical effects of, such as facilitated arousal, motivation, attention, memory and learning. The ligands have also been implicated in food-motivated behaviour. Further a relation with antipyretic activity has been disclosed.
Many different (synthetic) analogs of melanocortins have been . prepared and suggested to be of use in activating or blocking one or more of the MC-receptors. Z~his agonistic or antagonistic action has then been suggested to be useful in applications relating to pigmentation, nerve regeneration, etc.
. . _.,.
r.
~;,~,~~~,, .
Y' x ~.iWr . . 9. JUN. l 99 ~ i 5 ; C 1 4'LR OCfiR00I BUREAUY ;~~ i ~ 3342 73' 9 i NC'.
4?45 P. 18, 2 ,.
However, the melanocortins developed so far lack n .
specificity (selectivity) for the receptor* expr0 sed in the nsxvous system, and/or they lack ,i.n suffici~nt b ding .~ affinity or capability to,induce the receptor med at~d respoase or to block said response. Typically it ld ba desired. that a drug targeting the MC-raoeptors ba highly potent, orally adma.nistexabla, r~~asonalily resista~t to breakdaarn in the body (have a ~uf~Eicient half-lif~) cad able to cross the blood-brain barrier.
..
l0 The present invention provides peptides or peptid -like structures that meat the structural requirements o be useful as essentially 1tC-receptor specif;i.c drugs.
Thus th~ invention provides a pap~tida having spsc~.fic binding affinity for a melanocortino rec~ptor, preferabiylthe mc3, mc4 or mc5 recaptox conprisiag th~ sego~nve ~ .
X-Y--His-(D-2-Thienyl-Ale)-ar9-(Z) or x-x-His- ( 3-.pyridyi-D-Al~a ) -Arq- ( ~ ) whereby X and Y a~ amino acid residues and $ is Ian aromatic amino acid residue. , According to the prest~nt inv~entioxi the above po ides era aqonists for MsH aotivity ~rhfch as.~ highly potan. A vest' 25~ isportant contributioa to the high potency attributsd can to the 7-position (oountvd gut in t:h,~ oriqinsl -moiaculsr,) A
which should bs D-2--thianyl~Ala o~~ 3-pyridyi-D- a and for which only wty limited and very similar residue may be substituted without losi~xg the inc~raaso in aqonit potency.
AnothQr important contribut3.on to M88 agonistict3.Wity i*
the omission of residu~s 1-3 and/a~r the o~n3ssiono~ rasidusa 11-13. A hiqh.contribution to activity is also ovidad by p the pretence of an aromtic amino acid. residua at po ition~9.
positions 4 and 5 should normally not bs omitted these residuts should ba pras~nt though it is tar critical lass wdich aaa~no acid residues era preson~t at said itions. rt is clear that at least consarrvad substitutions era ilow*d for these positions, but also lass conserwd substitu tions grill 9. IorJ, 1999 s ~ ; 9 ~ ~~~~ ocfipo~~ ~ ~o~E~~Ty (3 i ) 3 ~422?39 ~ No. 4345 p. 19 3 ~ . .
histidine residue at position 6 .and the arginine esidve at position 8 ar~ quite important for activity and ~ ould only be replaced by very consorvatiw suJbstitutions, if a ~a11.
5~~ Especially the more important residues in general should not be. all replaced by substitutes i~z on~ and the sam~ molecule.
The preferred residue at position is Naftyi-alaai#ie, bo it. in the D-or is the L-configuration.
The presenc~ of this residue 1~ads to a fuxth~r i~ncreae~e i.r~
potency.
An a~n.W o acid residue at position 10 is not esse~ial. for the activity of the molecul~, but it does seem to have some effect. If a residue fs present j.t is pref~rred t,~at this residue is Qlycine or lysine, whe~rlby the latterihas the additional. advantage that it provides a reactive ;moiety whicb can be used to couple the peptids~ tv other molecules or to make the peptide cyclic. In the e~wnt that a cyc is paptida is to be produoed, which~is preferzed, since the ha f~lit~ of such a cyclic peptide is improved over the half-~ifo of the lin~ar fo~n, then a reactiv~s moiety at the other~end shouxd also be provided. Another advantage of having a ~yalic paptida is that these peptides tend to have a higher sal~axivity for MC-receptors, in particular an disulphide bridge'inareasee the selectivity for the MC-4 receptor. Cyclic peptid~s oan ho~nsyer also be produced by providing rea~"tivs moieties tside the essential sore that enable closury! of the loo ~uch ax p~ ~ ' reactive moieties leading to a lacaam.
The preferred residues it positions X and x, (nr~~ninq 4 and 5 in the original ~CTB-core) are mle for x and G1y! or lisp for 7r, whereby the presence of Asp loads to a further alivantage in having a raaatiw moiety for making a lactam.
The peptides.accordinq to the invention are gen~aliy more potent than irt~a itself. The prefQx~red peptides h~w poteneias of up to 100 timQS the potency of leg. Less pots t are within the scope of the present inwntioa, s ncelpoteency' ~ is not the only criterion which is required for ~a successful peptide-based drug. As already mentioned, half-l~~ife arid selectivity are also important par,~ters.
g~nerally not lead to'a significant loss of poten~y. The 9, dU~d. 1999 15; Ol ~'ER OCTR~JOIBUREAUX (3i ~ 334227319 . N0, 4345 P. 20 4 ~ , , . , . The tests used for~detes~aining the potency of thelpeptides are .
the regular in itro t~ate as well as an i vivo ~est i,n rats whereby the growing behaviour i;s measured: The.r~eults in a -groos~ng assay are good indications that the.pept~.des will be 5' able to activate the receptor and thereby the G-protein cascad~ coupled to said receptor and thus the pepltides can be used as agonists for ~sC-receptors . Targeting the ~.-receptor in particular of the MC..4 rec~pt«r fox which a t~cularly selective peptide has been provided by the pres invention in an agonistic manner is useful in the treatment of Ci~s-disorders, neurophat6ies, obesity, and in particuilar for diabetes related neuropathy, as 'sell as neuropat~y ~ a result of cytotoaic treatments (chemotherapy aird the if.ke ) .
Th~retore tho pres~nt invention also provides ph~rmscantical co~ositic~ns capable of treating the above conditions. Dosages for suoh treatments will usually be given once n~day in doses of about 1 erg to about 100 mg p~z~ dosage unit, preferably 10 erg - 10 mq, more preferably 5b~ uq - 1 mg. The~dosage should ZO zmsult is a concentration in the body of between!0,1 nay at~d 1 um. preferably 1 n~1 - 100 n~i, aaost preferably ~0 - 50 nl~l.
The compositions msy comprise the usual additivs~ for usual dosage formats for peptide drugs or for peptide ~leriv~sd drugs, The format is preferably an oral formulation :ua~ as s tablet, Z5 a granulate, a powder or a~liquid formulation, a~though enteral and parerrteral administrations may find ~ppiication as wail. particularly preferred are ~~o~positions~ wherein a peptide according to the xrrventio~a is combined w~.th a drug aiuiing to prevent or that leads t~~ neuropathy, s~ch as~insulin 3 0 arrd cytotoxio agents . ' The invention w~.ll be explained in spore detail the following eape~r3,m~enta3, part .
I
i 9. JU?u', 1999 13. ~2 VER OCTRO~aISLTREAUK (31; 3342273:9 NG, 4343 P, 2I
. .
i BXPBRII~~
Material sad ~tstbods ' l~le,Ianocortin Ifgand receptor activity 5 v guman embryonal kidney (8810 293) cells mere stabi~r transfected with the human MC3(Gantz et al. ~tBC 1993. 268a824~6-8250), human MC4 (Gaeta et al. 1993. JHC; 268a15174015179~) or human MC5 receptor constructs using the calcium ph4spha~t precipitation method. A6 a reporter plasmid 10 ~ ~ of the pCRE-LaGz vector (Chon tt al. 1995. Anal.Biochem. ZZ6~349-354), in 'vrhich a c~ responsiv! element drives ezprsssio~ of the Lacx gene, was transfected at ttu begj.aning of each ~~rioent. The day after pCRE~LacB transfection, cells were split in 96-wells plates.. After 48 hours cells were stimulated with var5ring concentrations of the MC recepto=~ ligands in ass~y miodium Dl~t t 0 :1 mg/ml HBA, 0 . i mi~~ ZBMx ) for 6 house . ' Cells were lysed in lysir buffer_by a freQae-thaw round, suk~stratQ buffer (60~! sodium phosphate, i mt~t MqClZ) 5 m!t ø-merea~tosthanol, 200 NQ/ml ONPG) was added and col.ls were incubat~rd at 37~C for 1 hour. The activation of the cAll~ signal~transd~ction pathway upon receptor activation was detected in a micro;plat~ reader (8forad le'del 3500) a 405nm using a colorimetri~ assay as dQecribed by Chen et a1.) Animals, 3aqpiantatson o! carrnctlar~, fntracerebrov$ritricti3ar injection .
dale ~Pistar rats woigt~ing 120-130 g were implant~d with cannulas into the foramen intrave~ntriculars uade~r hypno~o anaesthesia (8rakkee at al., Life 3cieace'vol. ih 19'79, ).
hats were allomsd to recover for .3 days and usodj for experiments during the neat 10 da;rs. In case that rats were used for more than one grooming e;~perimrat they ere allomsd to recover for at least 3 days between subsequo~itwxpe~riments, peptides (15 nq)~dissolvod in 3 N~~ saline (154 m~ NaCl) were injected i.c.v. by means of a Ha~~lton syring~s. 't3ruom,ing taste ' were performed according to (Gispc~n~ et al, Lab. ~llnim. Sci. 29 1975). Rats were placed into the obsorvatioa boos ima~ediately after the infection. obsezvaticn started 15 miniafter the i i CA 02275442 1999-06-16 j I
u. JUN, !9~'~ _~ 5: Q? UER UC'fROnBUREAUX i3!' 3~~4~~'l3' ~ 1VU, 4~?4'~
1.
fi , r .
injection and continued for 50 min. Grooming was cored each 15 sec over 50 min, thus the msxa.mal grooming scope for a rat is 200. ~ j _ 5~ syntbl,ia of peptides Pur'~f i ca -i on o ~~ ~ ' Preparative BpLC was carried out using a eaters P~sp 4000 .
liquid chromato9raph, eguipped wj.th a Waters aCM ~modui~ with two prepBak cartridges plus c~uardl cartridge ( 25:2~.0 mm) filled with Delta-Pak C18 material. Peptides tm~re detec~ad at 230 nm using a Waters 486 spectrophoton~a~ter with a prap~rativrr cell.
purifications wexe performrad in g~radfonts a*in9 v~ater with 0.11 trifiuoroacetic acid (TFA) and aaetonitrile.with O.lt TPA.
.1S i l~tettiods for synthesis and cyclisation of a-r~iSH p~ptides a ~ltinl~es~.id~ Svnthnw~t*
We .used a eanilton 1r11crolab 2200 to synthesise u~ to 40 peptides sinaltaneously at 30 ~1 scan. Thi 8a~iiton Microlab 2200 xaa programesd to delivor washing ~olveats and roagents to a rack with 40 individual 4 ml columsi* with filter, containing Rf.dk ( 4- ( 2 ~ , 4,' .-dinsthosypheny~-I s'nrx-aminos~hyl)-pheno:y) resin for peptide *ynthssf.~. The columns wets drained automatically after ~eaoh atop by vacuum. The 2S coupling Gyclo.rras based on Fmoc/»BTV (2-(1H-ben~otria~sol-1-yi)-1,1,3.,3-tetraa~thyZuronium hoaafluorophospha~e) chemistry [Fields ~t al.~ Peptide Research ~~, 95-101] usin~ double coupling steps of 40 minutes. Pep~tidos ire depr~tec~ted and cl~avod in 2 hre using 1.5 ml of as mixture o! tr~fiuoroacvtic acid/phenol/thioanisolelwater/ethanedithioi j 10/0.7510.5/0.5/0.25 and then precipitated twicelby adding hr~ane/disthy3.ether 111. The precipitate was dri~d and lyophi7.ised from water/acetoaitri7:e. , C~~~,n_ of n nt; ~. .
PLC purified' or crnd~e peptides wore used for cyriization. via a disulfide bridge with cysteiaes or via a lactajs br3,.dQo w3.th the side ehaias of aspartio acid and lysiaes~ !
A. disulfide bridge: crude MBJ06 (40 mg) was dissolved in 40 ml of a 0.5~ ammoniumbicarbonate~ solution at pH 8 and stirred overnight. After 24 hours no free sulphydryl groups were detected using Ellman's reagent and the product was lyophilized after addition of 0.5 ml of acetic acid. The y peptide was dissolved in 3 ml of 40~ acetic acid and purified by preparative HPLC in a gradient of 14$ to 21$ acetonitrile in water (containing 0.1~ TFA) i.n 30 min. Yield after purification: 22.8 mg.
B. Lactam bridge: a mixture of 20 ml of DMF (peptide grade), 26 mg of Benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP, 0.05 mmol) and 0.017 ml of DIEA
(0.1 mmol) was added to crude MH~J07 (20 mg, 0.012 mmol). The cyclization reaction was followed by analytical HPLC. After 2 . hours the mixture was acidified to pH 4 using 0.1 M HC1. The product was purified by preparative HPLC after dilution with 30 ml of water in a gradient of 21~ to 30~ acetonitrile in water, containing 0.1~ TFA, in 30 min. Yield 15.4 mg.
Results Screening of modified MSH peptides generated by PEPSCAN~
revealed several amino acids that increased MSH potency. D-2-thienyl-Ala and 3-pyridyl-D-Ala at position 7 of the MSH
peptide was the most potent contributors of increased MSH
potency. Naftyl-Ala at position 9 also increased MSH potency (figure 1b). Deletion of positions 1-3 and 11-13 further increased MSH activity. Figure 1 shows the dose-response curves for the following peptides:
MBU 23 *2GH6R7G# linear MBU 24 *2GH6R8G# linear MBU 27 *CGH6R8C# cyclic disulphate MBU 28 *2DH6R8K# cyclic lactam MBU 29 *2DH6R7K cyclic lactam * acetyl # carboxamid 2= Norleucine, 6= D-Thienyl, 7= 2-naftyl-L-alanine, 8= 2-naftyl-D-alanine 9. 1999 1;03 VER OCTROOIBUREAUX ('j1) JUN.3~4~~'~31~
' Nu, 434 r .
. 8 ( .
.
. , , (figures Positions 4~and 5 may not be critical 3 sand 4) -At position 9 an aromatic acid (1~,H,Y)is highly ~reterred figure 2 ) . Figures 3 to 9 show ithe ~ffect' of the' different -amina acid substitutions'.at positions4 to 10 of ale-I~cSS, 5' xespeatively.
. I
. induction The activity of these peptides can ref 9roodi.ng the behaviour ~ following intracer~hro~rentricular $tions is.
in jec shown in figure 6. 1500 nQ is thr~
lowest dose of iMBH that induces excessive grooming ~behav~Lour.The five cQmpouads tested hers are 100x pore potent than Ms8 in viva.
Table 1. Calculated 8C50 values f:or the three di~ferQnt cell lures . ' hHC3 hMC:4 hl~lC~
l~tJ 23 5.510-9 ~.~i,~0-9 a.dr i . 4.80-$
i~u 24 3.010-9 1.?v10-8 , i I~1 Z7 I.3I0-~ 6.G~10-g 2~.5a a-s MeU 28 6.310-7 1.710 8 4.5~0~8 i ' I~1 24 . 1.410-9 ~ 1.510-8 2.3~,Q~8 i a-MSH 1.410-a 2.610-8 .3.0x0'"$
i Legends ' 3o Pi~urs 1. Dose response curves ~:or the differsr~t ldSB peptides on eBlC 293 cells aabiy e=pressing husr~n MC3, KC~4 sad MC5 receptors. M8U 27 shows specificity for the HC4 ;receptor. 1~U
23 is a potent li9aad on all three receptor .
~ignre 1b. effect of the different aainoacids ~.t position 7 and 9 togeth~e= with MHtI 23 and IKB~J 24. All peptides wero tested at 100 ruK oa a8x 293 cells stabiy expreseinq the three different 1~C receptorsf MC3, MC4 ~~~d MC5 rtoieptc~rs. values i I
9. JUA, 19u ~ '~ ~, ; f?? E'R'R OCTR~JG i BURE~~n (3', i 334??"3~ ~ N0. 434 P, a ' ~ 9 '' ' i represent the percenteg~ activit~~ as cao~pared to the maximal acti~rity of 100 n1~1 8n1?-MBH.
' Eiguro ~ . 8ffect of sini~le aminoacid Bubstitutiqns at 5~ aminoacid position number.9 of t~P-MS8. On the x-p~cis the aminoacids replacing~the endogenous aminoacid (Try) are depicted. values represent~th~a percentage activity as cospared to thQ maximal activity of 10 nM at-MISH. All pept'~des were tested at 10 nlK on H81C 293 cells stably ~apress~g the. three different 1KC receptorsfi~C3,~ MC4 s.nd L~t05 receptors. at this position on an NDp-M8g background. an aromatic amino acid ie higtily preferred.
i~iqure 3. affect of single ami,noacid substitnti~tss at aminoacid position numbtr 4 of Nle-MSg. On the Xrtazis the aminoacid~t replacing the endogenous aminoacid (lit) err depicted. Values represent the pert~ntag~ activity as comparod to' the masimal activity , of 100 nl~!', a-M88. A11 peptides were tested at 100 aM oa 8810 293 cells etably eapress~ng the. three different NC receptors= i~C3, MC4 and' 1~C,5 receptors.
FiQnre ~. 8ffect of single aminoacid substitutilons at aaiinoacid position numtior 5 of l~Ie~1i88. On tho x~-asis the aminoacids replacing the endogenous aa~inoaaid (G~.u) are depicted. Values represent the percentage activity as cvosparetl to ttLe ma~cfaval activity of 100 nl! a-1~3H. All pep~ti.des were .
tasted at 100 n1~ on HBx 293 cells stably a:pressiing t1~ three diffQrent 1KC recaptorsi MC3, KC4 and MC5 rec~pto~rs.
Figure 5. 8tfect of singlo aminoacid substitutions at aminoaeid position number d of Nl~e-MBH. 0:1 the 7~-eels the as~inoacids replacing the endogenous aminoacid (bis) are depicted. valves represent the perc~ntagQ activ~:ty as compared to the ma:imsl activity of 100 nM a--MS8. rrll peptides mere tested at 100 nip op B8R 293 calls stably a=pressing the three different 1~C reoeptors=MC3, MC4 and- ICS receptors.
i q. JUIV, 1996 i~~:~3 VER ~.CTR;'~CrBURE"L''r 31; 3 ?""
r s 3~~ c r 319 ~ NC-, 435 F. 26 . 10 . ' Fignre '6. Effect of ~ single aminoacid substitutions at aminoacid poeitiou number 7 of N7.e-MSH. On the X-!axis the aminoacids replacing the endogenous aminoacid (Pte) are depicted. Values represent the perc~atage activity as coa~ared to th~ maximai'activity of 100 nr!t a-l~H. A11 pep#id~s were tested at l00 nli on eSE 293 call~i stably ezpress~lag the three different MC receptor:; MC3) MC4 and HCS racapto~s. ~ -Pfgure ~. Effect of single aminoacid substitution: at amfnoacid position n~ober 8 of Nl,e-1188. Ori the Z+axis the ~ami.noacids replacing the endogenous aminoacid (A~g) airs depiotrd. values represent tho percentage activity as comparod to the maximal activity of 100 n~!t a-M88. 7111 peptides arere tosted ~ at 100 ate on B~ 293 cell~~ stably expressing tlu~ three different MC receptors;MC3, MC4 and 1~C~ receptors. Only fiv~
aminoacid~substitutioria mere reefed.
Figure o. Hffect of single amincxcid substitutibris at aainoacid position nunber 9 of Nl.e~MSB. oa tho 3~aais the aminoacids replacing tho tr~dog~nous ~0.i.noacid (Tlrp) are dQpiotod. Valuos~repreetat the pa~rcantage activity as compared to the ma~zimal activity of 100 nl~t~ a-M88. 1111 ptpitides were tested at 100 nH on 881 293 cells aably eupross~iaQ tho tdrQe d~,ftorerit iIC-rlceptorap M~C3, KC4 and MC5 receptors.
, -Figure !, affect of single amincxoid substitutions act aoinoacid position numba,t 10 of 1~'1Q-MBH. 0n the~X-a:is t~
aminoacids replaci~aq the endogenous anninoacid ( (~ly ) are depictod. values represent the percentage activity as co~mparod to thc~ mazimel activity of 100 nil a-~tSH. 1111 ps~tides were ttst~d at 100 ~u~t ~ on 293 collr stably e:pres~ting the three different MC' receptors ~ ~IC3, Mc4 and xC5 receptors .
rigur~ lo. .
Peptide ~faductd groaioiag ~ .
' 3 ;ri saline, ~or 3 ;rl saline with ~eithor 15 ng 1~Q peptides or 190 ng a-l~lBH or 1500 ng a-~18s wau in jeoted i.c.v. in rats and grooming behaviour was snored (mean t s.d.).
Claims (12)
1. A peptide having MSH agonistic activity and having specific binding affinity for a melanocortine receptor, preferably the mc3, mc4 or mc5 receptor comprising the sequence X-Y-His-(D-2-Thienyl-Ala)-Arg-(Z) or X-Y-His-(3-pyridyl-D-Ala)-Arg-(Z) whereby X and Y are amino acid residues and Z is an aromatic amino acid residue.
2. A peptide according to claim 1 comprising the sequence X-Y-His-(D-2-Thienyl-Ala)-Arg-(2-naftyl-Ala) or X-Y-His-(3-pyridyl-D-Ala)-Arg-(2-naftyl-Ala) wherein X and Y are as defined in claim 1.
3. A peptide according to claim 1 or 2 comprising the sequence X-Y-His-(D-2-Thienyl-Ala)-Arg-(Z)-Z2 or X-Y-His-(3-pyridyl-D-Ala)-Arg-(Z)-Z2 wherein X, Y and Z are as defined in claim 1 or 2 and wherein Z2 is an amino acid residue.
4. A peptide according to claim 3 wherein Z2 is Gly or Lys.
5. A peptide according to any one of the aforegoing claims whereby Y is Gly or Asp.
6. A peptide according to anyone of the aforegoing claims wherein X is Nle.
7. A peptide according to any one of the aforegoing claims which is cyclic.
8. A peptide according to claim 7 whereby the cyclic peptide is produced by making an S-S bridge.
9. A peptide according to claim 7 whereby the cyclic peptide is produced by making a lactam.
10. A pharmaceutical composition comprising a peptide according to anyone of the aforegoing claims.
11. A composition according to claim 10, further comprising insulin or a functional equivalent thereof.
12. A composition according to claim 10, further comprising a cytotoxic agent.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AT96203567.1 | 1996-12-17 | ||
| EP96203567 | 1996-12-17 | ||
| PCT/NL1997/000703 WO1998027113A2 (en) | 1996-12-17 | 1997-12-17 | Melanocortin derivatives for specific binding of melanocortin receptor 3, 4 or 5 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2275442A1 true CA2275442A1 (en) | 1998-06-25 |
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ID=8224709
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002275442A Abandoned CA2275442A1 (en) | 1996-12-17 | 1997-12-17 | Melanocortin derivatives for specific binding of melanocortin receptor 3, 4 or 5 |
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| EP (1) | EP0946595A2 (en) |
| JP (1) | JP2001506996A (en) |
| CN (1) | CN1246868A (en) |
| AU (1) | AU5348098A (en) |
| CA (1) | CA2275442A1 (en) |
| WO (1) | WO1998027113A2 (en) |
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| ES2321162T3 (en) | 1997-04-15 | 2009-06-03 | Csir | PHARMACEUTICAL COMPOUNDS AND COMPOSITIONS THAT HAVE APPETIZING SUPPRESSIVE ACTIVITY. |
| IL145406A0 (en) | 1999-03-29 | 2002-06-30 | Procter & Gamble | Melanocortin receptor ligands |
| GB2355657B (en) | 1999-10-27 | 2004-07-28 | Phytopharm Plc | Inhibitors Of Gastric Acid Secretion |
| WO2001030808A1 (en) * | 1999-10-27 | 2001-05-03 | The Regents Of The University Of California | Methods and compounds for modulating melanocortin receptor-ligand binding |
| US6693165B2 (en) | 2000-01-18 | 2004-02-17 | Merck & Co., Inc. | Cyclic peptides as potent and selective melanocortin-4 receptor antagonists |
| GB0012370D0 (en) * | 2000-05-22 | 2000-07-12 | Quadrant Holdings Cambridge | Peptoids |
| GB2363985B (en) | 2000-06-30 | 2004-09-29 | Phytopharm Plc | Extracts,compounds & pharmaceutical compositions having anti-diabetic activity and their use |
| AU2002238106A1 (en) | 2001-02-13 | 2002-08-28 | Palatin Technologies, Inc. | Melanocortin metallopeptides for treatment of sexual dysfunction |
| CA2453515A1 (en) | 2001-07-11 | 2003-01-23 | Palatin Technologies, Inc. | Linear and cyclic melanocortin receptor-specific peptides |
| MXPA05011830A (en) | 2003-05-09 | 2006-01-26 | Novo Nordisk As | Peptides for use in treating obesity. |
| WO2005030797A2 (en) | 2003-09-30 | 2005-04-07 | Novo Nordisk A/S | Melanocortin receptor agonists |
| WO2005070468A2 (en) | 2004-01-21 | 2005-08-04 | Novo Nordisk A/S | Transglutaminase mediated conjugation of peptides |
| NZ590254A (en) | 2008-06-09 | 2012-07-27 | Palatin Technologies Inc | Melanocortin receptor-specific cyclic peptides for treatment of sexual dysfunction |
| EP2440572B1 (en) | 2009-06-08 | 2017-04-05 | Palatin Technologies, Inc. | Lactam-bridged melanocortin receptor-specific peptides |
| NZ596617A (en) | 2009-06-08 | 2014-04-30 | Palatin Technologies Inc | Melanocortin receptor-specific peptides |
| UY32690A (en) | 2009-06-08 | 2011-01-31 | Astrazeneca Ab | SPECIFIC PEPTIDES FOR MELANOCORTIN RECEPTORS |
| CN102725305B (en) | 2009-11-23 | 2016-08-24 | 帕拉丁科技公司 | Melanocortin-1 receptor-specific cyclic peptide |
| KR20120102716A (en) | 2009-11-23 | 2012-09-18 | 팔라틴 테크놀로지스 인코포레이티드 | Melanocortin-1 receptor-specific linear peptides |
| EP2513096A1 (en) | 2009-12-18 | 2012-10-24 | Janssen Pharmaceutica, N.V. | Substituted aminothiazolone indazoles as estrogen related receptor-a modulators |
| TW201130831A (en) | 2010-02-17 | 2011-09-16 | Janssen Pharmaceutica Nv | Aminothiazolones as estrogen related receptor-alpha modulators |
| AU2011218190A1 (en) | 2010-02-17 | 2012-09-06 | Janssen Pharmaceutica Nv | Aminothiazolones as estrogen related receptor-alpha modulators |
| EP2539364A1 (en) | 2010-02-26 | 2013-01-02 | Novo Nordisk A/S | Peptides for treatment of obesity |
| BR112012021231A2 (en) | 2010-02-26 | 2015-09-08 | Basf Plant Science Co Gmbh | method for enhancing plant yield, plant, construct, use of a construct, method for producing a transgenic plant, collectable parts of a plant, products derived from a plant, use of a nucleic acid and method for producing a product |
| WO2011149841A1 (en) | 2010-05-25 | 2011-12-01 | Janssen Pharmaceutica Nv | SUBSTITUTED THIAZOLIDINEDIONE INDAZOLES, INDOLES AND BENZOTRIAZOLES AS ESTROGEN-RELATED RECEPTOR-α MODULATORS |
| ES2538702T3 (en) | 2012-10-19 | 2015-06-23 | Txp Pharma Gmbh | Analogs of MSH-alpha and MSH-gamma |
| AU2015249540B2 (en) | 2014-04-22 | 2019-08-22 | Txp Pharma Gmbh | Peptide analogues with branched amino acid probe(s) |
| JP2020514365A (en) | 2017-03-15 | 2020-05-21 | ノヴォ ノルディスク アー／エス | Bicyclic compound capable of binding to melanocortin 4 receptor |
| US20210221867A1 (en) | 2018-05-15 | 2021-07-22 | Novo Nordisk A/S | Compounds Capable of Binding to Melanocortin 4 Receptor |
| WO2020053414A1 (en) | 2018-09-14 | 2020-03-19 | Novo Nordisk A/S | Bicyclic compounds capable of acting as melanocortin 4 receptor agonists |
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| WO1994022460A1 (en) * | 1993-04-05 | 1994-10-13 | University Patents, Inc. | Diagnosis and treatment of erectile dysfunction |
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1997
- 1997-12-17 CA CA002275442A patent/CA2275442A1/en not_active Abandoned
- 1997-12-17 AU AU53480/98A patent/AU5348098A/en not_active Abandoned
- 1997-12-17 JP JP52756598A patent/JP2001506996A/en active Pending
- 1997-12-17 CN CN97181332A patent/CN1246868A/en active Pending
- 1997-12-17 WO PCT/NL1997/000703 patent/WO1998027113A2/en not_active Application Discontinuation
- 1997-12-17 EP EP97950498A patent/EP0946595A2/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| WO1998027113A2 (en) | 1998-06-25 |
| WO1998027113A3 (en) | 1998-08-06 |
| EP0946595A2 (en) | 1999-10-06 |
| CN1246868A (en) | 2000-03-08 |
| AU5348098A (en) | 1998-07-15 |
| WO1998027113A8 (en) | 1999-09-02 |
| WO1998027113A9 (en) | 1999-11-04 |
| JP2001506996A (en) | 2001-05-29 |
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