GB2237571A - Gonadotropin releasing hormone analogues - Google Patents
Gonadotropin releasing hormone analogues Download PDFInfo
- Publication number
- GB2237571A GB2237571A GB9023748A GB9023748A GB2237571A GB 2237571 A GB2237571 A GB 2237571A GB 9023748 A GB9023748 A GB 9023748A GB 9023748 A GB9023748 A GB 9023748A GB 2237571 A GB2237571 A GB 2237571A
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- trp
- tyr
- ser
- pglu
- arg
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- 229940046085 endocrine therapy drug gonadotropin releasing hormone analogues Drugs 0.000 title abstract 2
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- 239000003488 releasing hormone Substances 0.000 description 1
- 230000027272 reproductive process Effects 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000003307 slaughter Methods 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000010009 steroidogenesis Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/23—Luteinising hormone-releasing hormone [LHRH]; Related peptides
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Endocrinology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
Gonadotropin releasing hormone analogues of formula: Pyroglu-His-Trp-Ser-A-B-C-D-Pro-E wherein A is His or Tyr; B is D-Arg, D-Lys, D-Trp, D-Ala, D-Nal (2) or D-(tBu) Ser; C is Leu or Trp; D is Arg, Gln, Tyr or Leu; E is Gly-NH2, D-Ala-NH2, NEt or azaGly provided that, when A is His, C is Leu, D is Arg and E is Gly-NH2, then B is other than D-Tyr.
Description
ANALOGUES OF GONADOTROPIN RELEASING HORMONE.
This invention relates to novel peptides and to their uses.
Gonadotropin releasing hormone (GnRH) is a decapeptide which
is secreted by the hypothalmic region of the brain.
Mammalian gonadotropin releasing hormone has the following
amino acid sequence p Glu- His- Trp- Ser- Tyr- Gly- Leu- Arg
Pro-Gly NH. There are however variant forms of mammalian
GnRH in other vertebrates, for example fish and birds.
The following is a table in which the primary structures of
GnRHs isolated from the brains of different vertebrates is
depicted.
Mammal PGlu-His-TrP-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2
Chicken I PGlu-His-TrP-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2
Salmon PGlu-His-TrP-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2
Chicken II PGlu-His-TrP-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2
GnRH and its analogues stimulate the release of pituitary
gonadotropin hormones and these in turn activate
gametogenesis and steroidogenesis in the gonads. Increased
release of gonadotropin hormones has been shown to stimulate
the reproductive process in animals and hence methods of
increasing the release of gonadotropin hormones has
application in the medical and agricultural spheres.
GnRH is also present in, and has effects on, the placenta, central nervous system, adrenals, gonads, and mammary tissue in mammals and hence GnRH analogies also have potential in medical and veterinary applications in regulating these tissues.
It is thus desirable that further and more effective analogues of GnRH be found to increase or decrease the release of gonadotropin hormones in various different classes of vertebrates.
Attempts have also been made to increase the gonadotropin releasing-activities of the various primary vertebrate GnRHs in mammals by incorporating D-amino acids into their structure.
In an article in Endocrinology, volume 124, pages 1830 to 1840, 1989 by J A King et al there is disclosed a chimaeric analogue of a naturally occurring vertebrate gonadotropin releasing hormone, pGlu-His-Trp-Ser-His-D-Tyr-Leu-Arg-Pro
Gly.NH2.
According to one aspect of the invention there is provided a peptide which is a chimaeric analogue of a naturally occurring vertebrate gonadotropin releasing hormone of the formula; pGlu-His-Trp-Ser-A-B-C-D-Pro-E wherein:
A is His or Tyr;
B is D-Arg, D-Lys, D-Trp, D-Ala, D-Tyr, D-Nal(2), or D-tertbut-Ser,
C is Leu or Trp;
D is Arg, Gln, Tyr, or Leu; and
E is Gly.NH2, D-Ala.NH2, NEt or aza-Gly, with the proviso that when A is His, C is Leu, D is Arg and
E is Gly.NH2 then B is not D Tyr.
By "chimaeric analogue" is meant a vertebrate gonadotropin releasing hormone wherein one or more of the amino acids at positions 5,7 or 8 are substituted with a different amino acid occurring in the corresponding position in another naturally occurring vertebrate gonadotropin releasing hormone.
Preferred peptides according to the invention include: pGlu - His - Trp - Ser - His - D - Trp - Leu - Arg - Pro
Gly NH2
I pGlu - His - Trp - Ser - Tyr - D - Trp - Trp - Arg - Pro
Gly NH2 II pGlu - His - Trp - Ser - His - D - Trp - Trp - Arg - Pro
Gly NH2 III pGlu - His - Trp - Ser - Tyr - D - Trp - Trp - Tyr - Pro
Gly NH2
IV pGlu - His - Trp - Ser - Tyr - D - Arg - Trp - Tyr - Pro
Gly NH2 V pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Tyr - Pro
Gly NH2 VI pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Gln - Pro
Gly NH2 VII pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Leu - Pro
Gly NH2 VII pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Arg - Pro
Gly NH2 IX pGlu - His - Trp - Ser - His - D - Arg - Trp - Arg - Pro
Gly NH2 X
In another aspect of the invention a peptide which is an analogue of a vertebrate gonadotropin releasing hormone having one of the following formulae is provided: pGlu - His - Trp - Ser - Tyr - D - Ala - Leu - Gln - Pro
Gly NH2 XIII pGlu - His - Trp - Ser - Tyr - D - Trp - Leu - Gln - Pro
NEt
XIV pGlu - His - Trp - Ser - Tyr - D - Arg - Trp - Leu - Pro
Gly NH2 XV pGlu - His - Trp - Ser - His - D - Trp - Trp - Tyr - Pro
Gly - NH2 XVI pGlu - His - Trp - Ser - His - D - Lys - Trp - Tyr - Pro
Gly NH2 XVII
The peptides of the invention have agonist properties relative to a naturally occurring vertebrate gonadotropin releasing hormone.
In yet another aspect of the invention there is provided the use of a peptide described above as a regulator for regulating the release of hormones in vertebrates and in particular the use of a peptide of any one the formulae IV,
VI, VIII, IX, XV, XVI or XVII to regulate the release of hormones in chickens.
Peptides of the invention are decapeptides having a sequence of ten amino acids. The first amino acid in the sequence occupies position one, the second amino acid in the sequence occupies position two and similarly for amino acids 3 to 10.
The invention provides a new set of GnRH analogues with altered activity compared with native mammalian GnRH of both mammalian and non-mammalian species. Some of the analogues of the invention are chimaeric GnRH analogues in which the amino acid at one of the variable positions along the ten amino acid sequence, namely positions 5, 7 or 8, is substituted with a different amino acid which occurs in the same position in another vertebrate GnRH. The chimaeric analogues of the invention further incorporate a D-amino acid at position 6 in their amino acid sequence. The chimaeric analogues may also be substituted at the carboxyl terminus with N-ethylamide (NEt) or D-Ala.NH2 or aza-Gly to enhance their activity. These are represented in Table 1.
The invention also provides a number of novel analogues of naturally occurring gonadotropin releasing hormones which include a D-amino acid in their amino acid sequence. One particular analogue of the invention has a further substitution in that Gly. NH2, which occupies position lo along- the amino acid sequence, is substituted with Nethylamide (NEt). These are represented in Table 2.
To delineate the functional importance of the variant amino acids in positions 5, 7 and 8, chimaeric analogues according to the invention were tested for luteinizing hormone (LH) and follicle stimulating hormone (FSH) releasing activities and for receptor binding activity in rat, sheep and chicken pituitaries.
The present study has investigated the activities of GnRH analogues incorporating combinations of the substitutions: His5, Trp7, Leu8, Gln8 and Tyr in mammalian GnRH together with the incorporation of D-Trp6, D-Lys6 and D-Arg6 on receptor binding and gonadotropin hormone release in sheep, rat and chicken pituitaries.
In the following examples the structure of each peptide is indicated by showing any substitution that it has in its amino acid sequence relative to the amino acid sequence of native mammalian gonadotropin releasing hormone. The position, in the amino acid sequence, of the substitution is indicated by a number in superscript. If no substitution is indicated in any particular position, such position is occupied by the amino acid, present in that position, in native mammalian gonadotropin releasing hormone. Therefore, for example, [Hiss - D - Trp6)GnRH represents an analogue of mammalian gonadotropin releasing hormone with the amino acid
Histidine in position 5 and the amino acid D - Tryptophan in position 6.
The peptides were prepared with an automated program on a
Beckman System 990 peptide synthesizer. Protected peptides were assembled by standard solid-phase methodology on p methylbenzhydrylamine-l% divinylbenzene-styrene copolymer resin (3, 4) utilising 50% trifluoroacetic acid ' in dichloromethane for N-terminal deprotection and dicyclohexylcarbodiimide for the coupling of the following
Boc-protected amino acids : Gly, Pro, Arg(Tos), Gln(Xan), Tyr(C12-Bzl), Lys(Cl-z), Leu.H20, Trp, D-Trp, Glu(OBzl),
His(Tos) and Ser(Bzl) as well as z-pGlu.
Peptides were cleaved from the resin and deprotected using 1% anisole in redistilled hydrogen fluoride (5). The peptideethylamide was cleaved with redistilled ethylamine and deprotected with hydrogen fluoride. The peptides were purified to greater than 96% homogeneity (based on the ratio of the integrated area under the main peak vs. the total integrated area recorded at 210 nm) by preparative C18 reversed-phase high performance liquid chromatography using a 0.25M triethylammonium phosphate pH 2,25/acetonitrile buffer system with desalting in a 0.1% trifluoroacetic acid/acetonitrile. Structures of peptides were confirmed by routine amino acid analysis.
Cultured chicken pituitary cell bioassay
Anterior pituitaries were removed from chicken heads (Golden
Grove Poultry Co., Cape Town) which had been kept on ice and washed with disinfectant ('Hibitane'; 0.5% chlorhexidine gluconate in 80% methanol), within 2 h of death. The pituitaries were collected into Dulbecco's Modified Eagles
Medium (Gibco, Grand Island, NY) buffered with 20 mM HEPES (pH 7.4) at room temperature, minced with a razor blade, and digested with collagenase for 1 h at 37 0C while undergoing continuous agitation by a slowly rotating stirrer.The collagenase solution consisted of 0.9% collagenase (w/v) (155
U/mg, Worthington Biochemical Corp., Freehold, NJ) and 18mg/l deoxyribonuclease (Miles Laboratories, Elkardt, IN) in HEPES
BSA buffer containing (mM): NaCl 137, KCl 5, Na2HPO4 0.7,
HEPES 25 (pH 7.2), Caul2 0.36, glucose 10 and 1% (w/v) BAS (fatty-acid free, Pentex fraction V, Miles Laboratories). At 10-min intervals the cells were triturated with a 5 ml pipette. The cell suspension was centrifuged twice at 500g for 5 min and the pellet resuspended each time in buffer A [NaCl 140mM, KCl 4mM, Na2HPO4 1.4 mM, glucose 8.3 mM, HEPES 20 mM (pH 7.4) and phenol red 6 mg/1] to which 0.5 mM EDTA and 0.3% (w/v) BSA was added.The cell suspension was then filtered through nylon gauze prior to dilution into
Dulbecco's Modified Eagles Medium with 10% foetal calf serum (Gibco), penicillin (60 mg/l), streptomycin (100 mg/l), neomycin (100 mg/l) and amphotericin B (20 mg/l) and dispensed into plastic tissue culture wells (Falcon, Oxnard,
CA) at a density of 1,3 pituitary equivalents per well (6well plates). One pituitary equivalent represents 5.0 to 6.7 x 105 cells. The cells were cultured at 37 0C in 5% CO2 for 24 h, after which the medium was replaced with fresh amphotericin B-free medium. Culture was then continued for a further 24 h.
Prior to stimulation, the cells were washed twice with buffer
A containing 1 mM CaCl2 and 0.1% BSA and pre-incubated twice for 5 min in the same buffer. Stimulations were carried out for 30 min at 370C in buffer A with 1 mM CaCl2 and 0.1% BSA.
The medium was collected, centrifuged and stored at -200C until radioimmunoassay of chicken LH. Mammalian GnRH was included in all bioassays as a reference standard for the comparative activity of the other GnRH analogues.
Cultured shee ',ituitarv cell bioassav
The biological activity of the synthetic GnRHs was assessed using cultured sheep anterior pituitary cells as previously described (1, 2). Anterior pituitaries were'dissected from sheep within 30 min of slaughter, diced and incubated in collagenase solution as described for chicken pituitaries.
The cells were resuspended in Minimum Essential Medium
Eagle (MEM) (with Hank's salts) (Gibco, Paisley, Scotland) containing 10% foetal calf serum (Gibco) and antibiotics and cultured (105 - 105/dish) at 370C in 5% CO2 for 4 or 5 days, with the medium being replaced after 3 days.
Prior to stimulation the cells were washed twice with MEM containing 10 % foetal calf serum and 4 times with serumfree MEM. Peptide stimulations (in triplicate) were conducted over 2 h in 1 ml serum-free MEM at 370C in 5% COz.
900 1 of medium from each test was collected into tubes containing 100 1 of HEPES-BSA buffer, centrifuged and stored at -200C until radioimmunoassay. Mammalian GnRH was included in all bioassays as a reference standard for the comparative activity of the other GnRH analogues.
Gonadotropin radioimmunoassavs
Ovine LH radioimmunoassays (RIA) were performed with duplicate aliquots of conditioned medium (or dilutions thereof) from stimulation experiments as previously described (6), except that the second antibody was linked to cellulose (Sac-Cel RD70, Wellcome Reagents Limited, Beckenham,
England). Purified ovine LH (LER- 1056-C2, gift from L.E.
Reichert) was iodinated by the chloramine-T method and free iodide was removed on a cellulose CF 11 (Whatman Inc.,
Clifton, NJ) column. A standard curve was generated with unlabelled sheep LH (NIH-LH-S18) and antiserum GDN 15 (gift from G.D. Niswender, Colorado State University) which had been raised against sheep LH.
Chicken LH was assayed as previously described (7), except that the second antibody was linked to cellulose (Sac-Cel
RD70).
Ovine FSH radioimmunoassays employed anti-oFSH-l antiserum (final dilution 1:160,000), radioiodinated oFSH-I-l and oFSH
RP-1 as standard. All reagents were supplied by S. Raiti (national Hormone and Pituitary Program, NIDDK) and utilised as instructed. Separation of bound and free 125I-oFSH was achieved with a second antibody conjugated to cellulose (Sac
Cel RD70). All comparative studies between analogues were performed in the same radioimmunoassay and the intra-assay coefficient of variation for radioimmunoassays were : 13.7% (chicken LH), 4.6% (sheep LH), 8.3% (sheep FSH).
Gonadotropin releasina hormone receptor bindina assavs The binding of synthetic vertebrate gonadotropin releasing hormones and analogues thereof to gonadotropin releasing hormone receptors in rat anterior pituitary membranes was investigated as previously described (1, 2). In the present study. a sheep pituitary gonadotropin releasing hormone receptor binding assay was also developed.
Anterior pituitaries from adult male rats (Long-Evans) or castrated adult male sheep were homogenised in ice-cold 10 mM
Tris-HC1 buffer, pH 7.4, containing 1 mM dithiothreitol, 1.0 mM EDTA and 0.1% BSA. The rat pituitary homogenate was centrifuged at 15,000 x g for 30 min at 40C to yield a membrane pellet which was resuspended in the Tris-HCl buffer.
The sheep pituitary homogenate, however, was centrifuged at 100 x g (5 min, 4 0C) and the supernatant recentrifuged at 3500 x g for 10 min at 4 0C. The resulting pellet was resuspended in the Tris-HCl buffer. The binding assay comprised incubation of the membrane preparation (0.16 rat or 0.05 sheep pituitary equivalents/tube), 60,000 cpm 125I-[D
Ala6-N Me-Leu7, Pro9-NHEt@GnRH [radioiodinated by a modification of the chloramine-T method (8) - specific activity by the self-displacement method, 1000-1600 Ci/ g) and increasing concentrations of the test peptides for 90 min at 40C. The assay was performed in glass test-tubes (12 x 75 mm) with a total volume of 0.5 ml 10 mM Tris-HCl buffer pH 7.4 containing 1 mM dithiothreitol, 1 mM EDTA and 0.1% BSA.
The incubation was terminated by dilution with 3 ml ice-cold phosphate buffered saline, pH 7.4, containing 1% BSA (PBS
BSA), followed by immediate vacuum filtration through glassfibre filters (GF/C; Whatman) presoaked in PBS-BSA. The filters were washed twice with 3 ml PBS-BSA, and the retained radioactivity counted. Non-specific binding was determined in the presence of 10-6 M unlabelled [D-Ala6, N Me-Leu7, Pro9
NET]GnRH and substracted from all samples (2% in rat and 26% in sheep).
Data reduction and potency estimates
ED50 values (concentrations required for half maximal gonadotropin release in bioassays or for half maximal inhibition of l25I-[D-Ala6-N MeLeu7, Pro9-NEt]GnRH binding) of
GnRH analogues were determined using a four-parameter, nonlinear curve fitting program (Allfit), forcing curves to share basal and maximal values. Kd values were calculated from the EDsos by the method of Cheng and Prusoff (9).
Standard errors of the mean were derived from Allfit estimates. Relative potencies were calculated from EDso and
Kd values relative to those for mammalian GnRH. All values presented are the means of several independent experiments.
Results
All of the analogues tested had enhanced LH-releasing activity in chicken pituitary cell cultures when compared with mammalian GnRH. Since mammalian GnRH and chicken GnRH
I in (the releasing hormone in the chicken hypothalamus) have similar activities, the analogues are also more active than chicken GnRH I. Analogues IV, VI, VIII, IX, XV, XVI, and
XVII had relative activities 500% or higher compared with mammalian GnRH and most of these are characterised by having
D-amino acid-Trp7-Tyr8 substitutions. In sheep pituitary cell cultures, analogues I, II, XIII and XIV display enhanced LHreleasing activity.
Receptor binding potency of the GnRH chimaeric analogues in sheep and rat pituitary was in general correlated with, LHreleasing potency in the sheep pituitary, however,two classes of analogues emerged in which the relative magnitude of these activities deviated significantly: 1 [His5-D-Trp6)GnRH exibited higher receptor binding activity than LH-releasing activity. (Table I) 2 The analogues with a Trp7-Tyr8 substitution characteristically displayed LH-releasing activity which is higher than their relative receptor binding potencies (Table
I).
Certain of the analogues, as indicated in Tables 1 and 2, showed potencies for FSH release which were similar to their potencies for LH release, in the sheep pituitary.
REFERENCES 1. Millar, R.P., and King, J.A. (1983) Endocrinoloov 113,
1364-1369 2. Millar, R.P., Milton, R.C.deL., Follett, B.K., and King,
J.A. (1986) Endocrinoloav 119, 224-231 3. Stewart, J.M., and Young, J.D. (1984) in Solid Phase Peptide Synthesis, 2nd edition, pp. 55-56, Pierce
Chemical Co., Rockford, Illinois 4. Merrifield, R.B. (1963) J. Am. Chem. Soc. 85, 2149
2154 5. Rivier, J., Kaiser, R., and Galyean, R. (1978) Biopolymers 17, 1927-1938 6. Millar, R.P., and Aehnelt, C. (1977) Endocrinoloov 101,
760-768 7. Follett, B.K., Scanes, C.G., and Cunningham, F.J. (1972)
J. Endocrinol. 52, 359-378 8. Nett, T.M., and Adams, T.E. (1977) Endocrinology
101,1135 - 1144 9.Cheng, Y.C. and Prusoff, W.H. (1973) Biochem. pharmacol.
22, 3099-3108 TABLE 1
The Estimated Potencies of Various Chimaeric Analogues of GnRH with regard to
Lutenizing Hormone (LH) Release and Receptor Binding Activity chimaeric GnRH Analcques LH RELEASE* RECEPTOR BINDING*
(chicken pituitary) (sheep pituitary) (sheep) (rat)
I. His5-D-Trp6 GnRH 3.5 4.2** 30.1 57.4
II. D-Trp6-Trp7 GnRH 3.1 7.6** 0.8 9.3
III. His5-D-Trp6-Trp7 GnRH 3.1 1.5**
IV D-Trp6-Trp7-Tyr8 GnRH 8.9 0.5** 0.01 0.25
V. D-Arg6-Trp7-Tyr8 GnRH 4.0 -
VI. D-Lys6-Trp7-Tyr8 GnRH 10.0 - 0.09
VII. D-Lys6-Trp7-Gln8 GnRH 2.5 - 0.07
VIII. D-Lys6-Trp7-Leu8 GnRH 6.7 0.1 0.02
IX. D-Lys6-Trp7 GnRH 5.0 2.0 1.1
X. His5-D-Arg6-Trp7 GnRH 2.1 1.0 0.8 *Potency relative to mammalian GnRH
GnRH refers to pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly NH2 **Similar potencies for FSH release were obtained TABLE 2
The Estimated Potencies of Various Chimaeric Analogues of GnRH with regard to
Lutenizing Hormone (LH) Release and Receptor Binding Activity
GnRH Analcques LH RELEASE* RECEPTOR BINDING*
(chicken pituitary) (sheep pituitaryl) (sheep) (rat)
CHICREN GnRH I Analoques
XIII. D-Ala6-Gln8 GnRH 2.1 2.6**
XIV D-Trp6-Gln8-Pro9 NEt GnRH 2.0 4.4** 1.3
SALMON GnRH ANALOGUES
XV D-Arg6-Trp7-Leu8 GnRH 5.4 0.09
CHICKEN GnRH II ANALOGUES
XVI His5-D-Trp6-Trp7-Tyr8 GnRH 6.9 0.4** 0.02 0.99
XVII His5-D-Lys6-Trp7-Tyr8 GnRH 5.0 0.5 0.2 *Potency relative to mammalian GnRH
GnRH refers to pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly NH2 **Similar potencies for FSH release were obtained
Claims (9)
- ChAItIS: 1. A peptide which is a chimaeric analogue of a naturally occurring vertebrate gonadotropin releasing hormone of the formula pGlu-His-Trp-Ser-A-B-C-D-Pro-E wherein A is His or Tyr; B is. D-Arg, D-Lys, D-Trp, D-Ala, D-Tyr, D-Nal(2) or D tert-but-Ser; C is Leu or Trp; D is Arg, Gln, Tyr or Leu; and E is Gly.NH2, D-Ala.NH2, NEt or aza-Gly, with the proviso that when A is His, C is Leu, D is Arg and E is Gly.NH2 then B is not D-Tyr.
- 2. A peptide according to claim 1 which is: pGlu - His - Trp - Ser - His - D - Trp - Leu - Arg - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - Trp - Trp - Arg - Pro Gly NH2; pGlu - His - Trp - Ser - His - D - Trp - Trp - Arg - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - Trp - Trp - Tyr - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - Arg - Trp - Tyr - Pro Gly NHz; pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Tyr - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Gln - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - - Lys - Trp - Leu - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - Lys - Trp - Arg - Pro Gly NH2; or pGlu - His - Trp - Ser - His - D - Arg - Trp - Arg - Pro Gly NHz;
- 3.A peptide which is an analogue of a vertebrate gonadotropin releasing hormone which is: pGlu - His - Trp - Ser - Tyr - D - Ala - Leu - Gln - Pro Gly NH2; pGlu - His - Trp - Ser - Tyr - D - Trp - Leu - Gln - Pro NEt; pGlu - His - Trp - Ser - Tyr - D - Arg - Trp - Leu - Pro Gly NH2; pGlu - His - Trp - Ser - His - D - Trp - Trp - Tyr - Pro Gly - NH2; or pGlu - His - Trp - Ser - His - D - Lys - Trp - Tyr - Pro Gly NH2
- 4. A peptide according to claim 1 for use in a method of regulating the release of hormones in vertebrates.
- 5. A peptide according to claim 2 for use in a method of regulating the release of hormones in vertebrates.
- 6. A peptide according to claim 3 for use in a method of regulating the release of hormones in vertebrates.
- 7. A pharmaceutical composition for use in the regulation of the release of hormones in vertebrates, which comprises at least one peptide as claimed in any one of claims 1 to 6.
- 8. A peptide which is a chimaeric analogue of a naturally occurring vertebrate gonadotropin releasing hormone are substantially as hereinbefore described.
- 9. A peptide which is a chimaeric analogue of a naturally occurring vertebrate gonadotropin releasing hormone. and substantially as hereinbefore described with reference to the examples.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ZA898317 | 1989-11-01 |
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| Publication Number | Publication Date |
|---|---|
| GB9023748D0 GB9023748D0 (en) | 1990-12-12 |
| GB2237571A true GB2237571A (en) | 1991-05-08 |
| GB2237571B GB2237571B (en) | 1993-10-20 |
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ID=25579889
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|---|---|---|---|
| GB9023748A Expired - Fee Related GB2237571B (en) | 1989-11-01 | 1990-11-01 | Analogues of gonadotropin releasing hormone |
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| WO2001074377A1 (en) * | 2000-03-31 | 2001-10-11 | Siler Khodr Theresa M | Non-mammalian gnrh analogs and uses thereof in tumor cell growth regulation and cancer therapy |
| WO2003093304A1 (en) * | 2002-05-03 | 2003-11-13 | Neurogenex Co., Ltd. | Agonists and antagonists of gonadotropin-releasing hormone-2, and use thereof |
| WO2003016331A3 (en) * | 2001-08-17 | 2003-11-20 | Theresa M Siler-Khodr | Non-mammalian gnrh analogs and uses thereof in regulation of fertility and pregnancy |
| WO2003020205A3 (en) * | 2001-08-28 | 2004-01-08 | Theresa M Siler-Khodor | Non-mammalian gnrh analogs and uses thereof in regulation of fertility and pregnancy |
| WO2005018657A3 (en) * | 2003-08-12 | 2005-04-07 | Theresa M Siler-Khodr | NON-MAMMALIAN GnRH ANALOGS AND USES THEREOF IN REGULATION OF FERTILITY AND PREGNANCY |
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| EP3571213A1 (en) * | 2017-01-20 | 2019-11-27 | Immune System Regulation Holding AB | Novel compounds (immunorhelins) |
| US11304990B2 (en) | 2017-01-20 | 2022-04-19 | ISR Immune System Regulation Holding AB (publ) | Use of known compounds—intracellular infections |
| US11672842B2 (en) | 2017-02-22 | 2023-06-13 | ISR Immune System Regulation Holding AB (publ) | Gonadotropin-releasing hormones for use as adjuvant immunotherapeutics |
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| GB2228262A (en) * | 1989-01-25 | 1990-08-22 | Nat Inst Immunology | Antigenic derivative of GnRH |
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- 1990-11-01 GB GB9023748A patent/GB2237571B/en not_active Expired - Fee Related
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| US4005063A (en) * | 1973-10-11 | 1977-01-25 | Abbott Laboratories | [Des-gly]10 -GnRH nonapeptide anide analogs in position 6 having ovulation-inducing activity |
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| US4618598A (en) * | 1982-04-12 | 1986-10-21 | Duke University | Method of regulating hormone function or release |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001028576A3 (en) * | 1999-10-15 | 2001-09-13 | Siler Khodr Theresa M | NON-MAMMALIAN GnRH ANALOGS AND USES THEREOF IN REGULATION OF FERTILITY AND PREGNANCY |
| WO2001074377A1 (en) * | 2000-03-31 | 2001-10-11 | Siler Khodr Theresa M | Non-mammalian gnrh analogs and uses thereof in tumor cell growth regulation and cancer therapy |
| US7834141B1 (en) * | 2000-03-31 | 2010-11-16 | Theresa Siler-Khodr | Non-mammalian GnRH analogs and uses thereof in tumor cell growth regulation and cancer therapy |
| WO2003016331A3 (en) * | 2001-08-17 | 2003-11-20 | Theresa M Siler-Khodr | Non-mammalian gnrh analogs and uses thereof in regulation of fertility and pregnancy |
| WO2003020205A3 (en) * | 2001-08-28 | 2004-01-08 | Theresa M Siler-Khodor | Non-mammalian gnrh analogs and uses thereof in regulation of fertility and pregnancy |
| WO2003093304A1 (en) * | 2002-05-03 | 2003-11-13 | Neurogenex Co., Ltd. | Agonists and antagonists of gonadotropin-releasing hormone-2, and use thereof |
| KR100808921B1 (en) * | 2002-05-03 | 2008-03-03 | 주식회사 뉴젝스 | Agonists and Antagonists of Gonadotropin-releasing Hormone-2 and Their Uses |
| WO2005018657A3 (en) * | 2003-08-12 | 2005-04-07 | Theresa M Siler-Khodr | NON-MAMMALIAN GnRH ANALOGS AND USES THEREOF IN REGULATION OF FERTILITY AND PREGNANCY |
| US9388216B2 (en) | 2006-06-16 | 2016-07-12 | Medical Research Council | Methods of treating reproductive cancer |
| US8536124B2 (en) | 2010-03-19 | 2013-09-17 | National Pingtung University Of Science & Technology | Artificial decapeptide for inducing vitellogenesis in fish |
| WO2018134373A1 (en) * | 2017-01-20 | 2018-07-26 | Immune System Regulation Holding Ab | Novel compounds (immunorhelins- intracellular infections) |
| CN110431147A (en) * | 2017-01-20 | 2019-11-08 | 免疫系统调节控股有限公司 | Noval chemical compound (immunostimulatory peptides-intracellular infection) |
| EP3571213A1 (en) * | 2017-01-20 | 2019-11-27 | Immune System Regulation Holding AB | Novel compounds (immunorhelins) |
| JP2020505378A (en) * | 2017-01-20 | 2020-02-20 | イミューン システム レギュレェイション ホールディング エービー | New compound (immunoherin-intracellular infection) |
| US11304990B2 (en) | 2017-01-20 | 2022-04-19 | ISR Immune System Regulation Holding AB (publ) | Use of known compounds—intracellular infections |
| US11434258B2 (en) | 2017-01-20 | 2022-09-06 | ISR Immune System Regulation Holding AB | Compounds (immunorhelins) |
| JP2022153565A (en) * | 2017-01-20 | 2022-10-12 | アイエスアール イミューン システム レギュレイション ホールディング アクチエボラグ（パブル） | Novel compounds (immunorhelins-intracellular infections) |
| US11564969B2 (en) | 2017-01-20 | 2023-01-31 | ISR Immune System Regulation Holding AB (publ) | Immunorhelin compounds for intracellular infections |
| AU2018209239B2 (en) * | 2017-01-20 | 2023-02-02 | ISR Immune System Regulation Holding AB (publ) | Novel compounds (immunorhelins- intracellular infections) |
| US11672842B2 (en) | 2017-02-22 | 2023-06-13 | ISR Immune System Regulation Holding AB (publ) | Gonadotropin-releasing hormones for use as adjuvant immunotherapeutics |
Also Published As
| Publication number | Publication date |
|---|---|
| GB9023748D0 (en) | 1990-12-12 |
| CA2029018A1 (en) | 1991-05-02 |
| GB2237571B (en) | 1993-10-20 |
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| Date | Code | Title | Description |
|---|---|---|---|
| PCNP | Patent ceased through non-payment of renewal fee |