HYPOGLYCAEMIC PEPTIDES

Abstract: A compound of general formula (I), wherein X is hydrogen, -CHCONH2 or -CH2CH2CONH2; each of R1, R2 and R3 is an L- alpha -amino acid, a delta -amino acid or an epsilon -amino acid; R4 is an L or D alpha -amino acid, a delta -amino acid or an epsilon -amino acid; and each of R5 to R11 is hydrogen or an L or D alpha -amino acid, a delta -amino acid or an epsilon -amino acid, or a pharmaceutically acceptable salt thereof. Methods of synthesis of peptides according to general formula (I), compositions containing the peptides, and uses of the peptides are also disclosed and claimed.

Description: HYPOGLYCAEMIC PEPTIDES This invention relates to methods of controlling blood glucose levels, and in particular to novel compounds with hypoglycaemic activity and to processes for producing these compounds. Background of the Invention The principal factor controlling blood glucose levels is the hormone insulin. Diabetes may result either from insufficient production of insulin or from resistance to the action of circulating insulin. su B sτrruτE SHEET It has been suggested that a peptide fragment of human growth hormone (hGH) , derived from enzymic digestion of pituitary, can counter the effects of natural inhibitors of insulin (Australian patent application no. 87961/75 by Choay S.A. ). This (^.-peptide chain, having sequence leu-ser-arg-leu-phe-asp-asn-ala, is comprised of amino acids all of which have the L configuration. This peptide, equivalent to hGH 6-13, was stated to be able to counteract the inhibition of insulin by somantin, a degradation product of growth hormone, and was thought to potentiate the effects of insulin, to induce synthesis of insulin receptor sites on cells, and to sensitize β> cells in the islets of Langerhans to glucose. However, it has subsequently been found that the peptide disclosed does not have these activities. In fact, inconsistency in demonstrating the claimed biological activities, due to the absence of detailed knowledge of the precise conformational structure of the active substance, prevented further development and validation of the original work by Bornsten and co-workers, inventors in respect of 879661/75, and this application lapsed. We have now prepared a family of novel j>-i ide linked L-aspartyl-L-asparaginyl peptides wherein the general structure of the peptide may additionally involve between 2 and 6 other o.-amino acid residues of either the D- or L- stereochemical form, or involve other non Λ-amino acid substitutions at positions 1 or 6 to 12. Furthermore, the in vitro and in vivo properties of these yS-imido-L-aspartyl-L-asparaginyl peptides as potent hypoglycaemic agents with functional capabilities of potentiating insulin action are demonstrated. In this application we describe (i) the synthesis of these peptides via solid and solution phase peptide synthetic protocols ? (ii) the purification and structural characterization of these peptides; (iii) the in vitro evaluation of these peptides in various biological assay systems; (iv) the evaluation of these peptides in lowering blood glucose levels in vivo. Brief Description of the Figures Figure 1 represents a stereochemical view of a representative peptide according to the invention; Figure 2 represents far ultraviolet circular dichroic spectra for synthetic peptide Pep (6-13) A -aspartimido form I(_) and synthetic peptide Pep (6-13) λ-form ( ). Spectra recorded in water at 21°C using 1 mg/ml solutions in 0.1 mm cells; Figure 3 represents infra-red spectrum for active synthetic peptide (6-13) -aspartimido form I. The spectrum was measured between 2000-1500 cm after the peptide sample (ca. 5 mg) was pressed into a KBr disc; Figure 4 represents infra-red spectrum for synthetic peptide Pep (6-13) pC-form, inactive. The spectra were measured between 200-1500cm . (a) the sample (ca. 5 mg) of peptide was pressed into a KBr disc, (b) the sample (ca. 5 mg) of peptide was saturated with Nujol to form a mull; Figure 5 represents fast atom bombardment (FAB) mass spectrometric spectra of the synthetic peptides active A -aspartimido form I Pep (6-13) and inactive oi -form Pep (6-13); Figure 6 represents H N.M.R. spectrum of active A -aspartimido form I synthetic peptide Pep (6-13); Figure 7 represents paper electrophoresis comparing synthetic peptide (ϊ -aspartimido form I Pep (6-13) active and synthetic peptide o(-form dissolved in water and dilute (1M) NaOH. (a) Electrophoresis was performed at pH 6.5 for 1 h at 1.5 kV. (b) Electrophoresis was performed at pH 3.0 for 2.5 h at 1.5 kV. Peptides were detected by spraying with ninhydrin; Figure 8 represents HPLC chromatograms or synthetic Pep (6-13) peptides using a shallow gradient (0% - 20% Solvent B over 20 min) . (a) Injection of purified synthetic Pep (6-13) A -aspartimido form I, active. (b) Injection of partly hydrolysed synthetic Pep (6-13) /£ -aspartimido form I. (c) Simultaneous injection of inactive synthetic Pep (6-13) oi -form peptide and active fi -aspartimido form I Pep (6-13) peptide Λ _ form; Figure 9 represents the change in blood glucose level in rats following injection of various synthetic inactive orm and active IL-aspartimido forms of peptides according to the invention; Figure 10 represents the effect of various doses of Pep (6-13) k imide during intravenous insulin sensitivity tests on normal rats; Figure 11 represents the effect of Pep (6-13) on glucose tolerance tests in normal rats; Figure 12 represents in vivo testing of Pep (6-13) (Asn 11 Asp 12 ) (Figure 11a) and Pep 6-13 (Asp 12 ) (Figure lib) for insulin-potentiating activity on "untrained" female mice. The dotted lines represent control animals (injected with 0.2 ml of saline) in each case. The unbroken lines represent test animals (injected with 2.4 mg/kg of peptide in 0.2 ml of saline) . Insulin was injected at t ; Figure 13 represents in vivo testing of peptides for insulin-potentiating activity on "untrained" and "trained" male mice. The dotted line represents the mean fall in blood glucose for control animals (injected with 0.2 ml of saline). The unbroken line represents the mean fall in blood glucose for test animals, injected with 2.3 mg/kg of peptide in 0.2 ml of saline, except for (a), in which the dose was 2.4 mg/kg. (a) Pep 6-13 (Gln 11 Asp 12 ) , "untrained" (b) Pep 6-13 (Gln 11 Asp 12 ) , "trained" (c) Pep 6-13 (Asn 1:L Asp 12 ), "trained" (d) Pep 6-13 (Asp 12 ), "trained" Insulin was injected at t ; Figure 14 represents the in vitro effect of Pep (6-13)4imide on glucose oxidation in isolated rat adipocytes in the absence and in the presence of various levels of insulin. Summary of the Invention According to one aspect of the present invention there is provided a compound of general formula I: O I R 10 ~ R 9 ~ R 8 " R 7 ~ R 6 ~ R 5 ~ R 4 H — N » — ' • CHlCOR 3 - R ~ R ι O 12 11 10 9 8 7 6 5 4 2 2 1 Position number wherein X is hydrogen, -CHCONH- or -CH 2 CH 2 CONH 2 ; each of R,, _ and R-, is an L- o. -amino acid, a o -amino acid or an £-amino acid; R 4 is an L or D ø-amino acid, a 5-amino acid or an £-amino acid; and each of R_ to R-,-, is hydrogen or an L or D o^-amino acid, a 5-amino acid or an £-amino acid, or a pharmaceutically acceptable salt thereof. Preferably R_ is a L or D o<--amino acid, a o-amino acid or an -amino acid. Preferably each of R,, R- and R, is selected from the group consisting of alanine, glycine and phenylalanine. Preferably R. is a hydrophobic amino acid; more preferably R. is selected from the group consisting of L-phenylalanine, L-tyrosine, L-tryptophan and L-histidine. • Preferably R 5 is a hydrophobic amino acid; more preferably R^ is selected from the group consisting of L- or D- leucine, isoleucine and histidine. _ R fi is a basic amino acid, and is preferably selected from the group consisting of L- or D- arginine, lysine, and histidine. R-, is preferably a substantially hydrophobic amino acid, and is preferably selected from the group consisting of L- or D- serine, leucine and isoleucine. 8 is preferably selected from the group consisting of L- or D- leucine, phenylalanine, proline or isoleucine, an £ -amino acid such as 6-aminohexanoic acid, 4-aminocyclohexane-l-carboxylic acid or similar amino acids. Each of Rg and R-, - is preferably a substantially hydrophobic amino acid, and is most preferably selected from the group consisting 'of L- or D- leucine, phenylalanine, proline, isoleucine or tyrosine. Rg, R Q or R-. Q may optionally be coupled to R-, via a bifunctional agent such as 6-aminohexanoic acid. The amino acids may optionally have side chain substituents. • - R l' R 2' R 8' R 9' or R 10 ma ^ k e absent from the peptide, allowing coupling between C-terminal and N-terminal amino acids via a bifunctional agent such as 6-aminohexanoic acid. Rg is equivalent in terms of sequence alignment to residue 8 of human growth hormone. The cyclic imide structure as shown in general Formula I is essential for activity. Activity increases with chain length up to 8 residues. A stereochemical view of a representative peptide is shown in Figure 1. The peptide according to the invention forms an amphipathic helix structure having a fluid hydrophobic face and a second face carrying an exposed imide group. It will be apparent to persons skilled in the art that pharmacologically active analogues and derivatives of the compound of general formula I are within the scope of the invention, e.g. o-, m-, or p-aminobenzoic acid or af- or o -lactams in place of the aspartimide ring. According to a second aspect of the invention there is provided a method of solid phase synthesis of a peptide of general formula I comprising protection of the e4-amino functionality of added amino acids using t-BOC or f-MOC and treating the resin-bound protected j< /straight chain L-aspartyl-L-asparaginyl peptide with a base to produce the stable i-imido form of the peptide. The base is a dialkylamine such as piperidine, or a trialkylamine, preferably triethylamine. According to a third aspect of the invention there is provided a method of lowering the level of blood glucose in a mammal in need of such treatment, comprising administering to that animal a hypoglycaemically effective dose of a compound of general formula I. Said compound may optionally be administered together with or in conjunction with a second hypoglycaemic agent. Detailed Description of the Invention The invention will now be described in detail by way of example only, with reference to the accompanying drawings. JL. PEPTIDE SYNTHESIS The various peptide analogues of the general formula I above were synthesised either by standard solution phase synthetic procedures or by using modifications of the Merrifield solid phase peptide synthesis procedure (R.B. Merrifield, J. Amer. Chem. Soc, 8_5, 2149-2145, 1963), of which the following protocols are representative. Peptides were initially synthesised in the alpha/straight chain form with all amino acid side chain groups protected. For the t-BOC-synthetic approach, the o -amino functionality of the incoming amino acid was protected with the tert-butyloxycarbonyl (t-BOC) group with deprotection, following coupling, with trifluoroacetic acid in CH 2 C1 2 . Side chain protecting groups such as O-benzyl, tosylbenzylester and chlorobenzyloxycarbonyl were used for the trifunctional amino acids. The initial coupling to the benzhydrylamine resin employed a 3-fold excess of the designated side chain protected amino acid R, , dissolved in N,N -dimethylformamide (DMF) . Subsequent couplings of the second, third, eighth amino acid (counting asp and asn separately) employed a 2 fold excess of side chain protected amino acid in DMF. The amino acids were sequentially activated using tert-butanol and dieye1ohexylcarbodiimide in approximately 5ml DMF and allowing reactants to stir at room temperature for 5-10 min. The acylurea was removed by filtration and the asymmetrical anhydride added to the resin. The suspended resin was stirred at room temperature for reaction times generally between 60 and 120 min. Each coupling was monitored for unreacted amino groups using 2,4, 6-trinitrobenzenesulphonic acid. Similar synthetic strategies with the symmetrical protected amino acid anhydrides or the pentachlorophenylesters led to the preparation of identical products but in different yields. After the final coupling reaction was completed, the fully protected resin-bound peptide was treated with a reagent grade trialkylamine such as triethylamine (15- 20 ml per g resin) in order to convert the L-aspartyl-L-asparaginyl moiety to the stable A-imido form. The resin-bound peptide was side chain deprotected with trifluoroacetic acid, and the deprotected (L-> -imido-L-aspartyl-L-asparaginyl peptide analogue cleaved from the resin using liquid hydrogen fluoride as the acid and dimethylsulphide/p-cresol or thioanisole as the scavenger. Reaction times were between 30 and 120 min at 0°C. Workup following cleavage involved standard procedures of peptide synthesis. The deprotected peptide was removed from the resin by washing with trifluoroacetic acid, then water, then water/acetonitrile mixtures. The washes were then combined and lyophilised. The crude dried product was chromatographed by anion exchange chromatography on Whatman DE 52 and then desalted on Biogel P2 gels . Further purification was achieved using reversed phase HPLC with octadecylsilica as stationary phase and gradients of 0-75% water-acetonitrile - 0.1% trifluoroacetic acid as mobile phase. A similar synthetic strategy using f-MOC ( 9-fluorenylmethyloxycarbonyl group) protection has also been employed. The synthetic peptides were characterised by amino acid compositional analysis, analytical reversed phase HPLC with several elution systems, fast atom bombardment mass spectroscopy (FAB-MS) , paper electrophoresis at pHδ.O, FTIR-infrared spectroscopy, aminopeptidase M cleavage, H -NMR spectroscopy and circular dichroism (CD) spectroscopy. It was found that the synthesis protocol could be manipulated in order to produce all c or all imide form of the peptide. In the absence of alkylamine treatment, all was formed, and vice versa. Representative results from these characterizations are illustrated for the peptide leu-ser-arg-leu-phe-asp-asn-ala ( c -form, inactive), and for the peptide leu-ser-arg-leu-phe-Λ -imidoasp-asn-ala ( S -imido form I, active) . Table 1 shows representative results of amino acid composition analysis following acid hydrolysis and aminopeptidase M cleavage. TABLE 1 Amino Acid Leu Ser Arg Leu Phe Asp Asn Ala Sequence 0^—form Acid Hydrolysis' 1.03 0.78 0.93 1.03 0.98 0.98 0.98 0.96 Enzymic Hydrolysis 1.02 0.98 1.02 0.98 0.98 0.92 1.00 aspartimide Acid Hydrolysis 3 1.02 0.86 0.86 1.02 0.97 0.99 0.99 1.00 Enzymic . b Hydrolysis 1.06 0.99 1.06 1.00 a) 24 h acid hydrolysis at 110 U C in 6M HCl. b) 24 h aminopeptidase M digestion. Results of circular dichroism spectroscopy, infra-red spectroscopy, FAB-MS, and H-NMR spectroscopy are illustrated in Figures 2 to 6 respectively. Without wishing to be bound by any proposed mechanism for the observed effects, it appears that the o form is in an extended configuration, while the -aspartimido form I has one completely hydrophobic face, with the β -imido group at 90° to this hydrophobic face. This conclusion is in accord with results obtained from correlation Overhauser spectroscopy (COSY) and nuclear Overhauser enhanced spectroscopy (NOESY) NMR spectroscopy for all cases of the representative active f> -imide analogues. Figure 8 shows results of reversed phase HPLC on ODS-silica with shallow gradients of 0.1% trifluoracetic acid - acetonitrile-water. 2_. BIOLOGICAL ACTIVITIES Considering the physiology of hypoglycaemia and the possible molecular actions of peptide analogues with hypoglycaemic and insulin potentiating properties, the following in vitro and in vivo assays were chosen to assess the potency of the various synthetic Λ -imido-L-aspartyl- L-asparaginyl peptide analogues. (a) In vitro bioassays (i) (ii) (iii) Glucose uptake In these assays collagenase-dispersed rat adipocytes, cultured muscle cells (L6 rat skeletal muscle cell line) and hemidiaphragm preparations were employed. (b) In vivo bioassays (i) Effect on basal blood glucose levels (ii) Effect on insulin release (iii) Intravenous insulin tolerance test (iv) Activation of glycogen synthase Representative data on the biological properties of the active /^-imidoaspartylasparaginyl form I peptides and their inactive Λ -form peptide analogues are presented. Table 2 shows the decrease in blood glucose level in rats following intravenous injection of peptides according to the invention. Injection of the pt-form had no significant effect; however, injection of the A>imide form of Pep (6-13) or Pep (6-15) resulted in a significant hypoglycaemic effect. TABLE 2 Peptide Cone, of No. of Decrease Difference 2P Value Peptide animals in blood from control t—test j-imol/lOOg from basal mean Values are changes (means +_ SEM) of blood glucose from the basal level (4.0 - 4.4 mmol glucose/1) at time 0. Differences between the control and test groups were statistically evaluated according to Student's t-test. N.S. indicates 2P ~ > 0.05. Similar results are illustrated graphically in Figure 7. The <* form of Pep (6-13) was inactive for several variants tested (Ala 11 , Gly 10 , Gly 10 Pro i:L , Pro 11 , Tyr 10 ) whereas the f» imide form was always active. Similarly Pep (6-11), Pep (6-12), Pep (6-13), Pep (6-15) were active only in the A imide form. However, for Pep (9-13) neither the K. form nor the Λ imide form was active in this test. The Λ imide form of peptides according to the invention was also found to potentiate the action of insulin during intravenous insulin sensitivity tests carried out on rats trained to eat on a scheduled time basis. Rats were anaesthetised with pentobarbitone, and blood samples taken at various times after intravenous infusion of insulin. Control animals were untreated or were given insulin only; test animals received insulin plus the /> imide form of Pep (6-13) . The results are shown in Figure 10. The effect depended on the dose of peptide. Figure 11 shows that Pep (6-13) {i imide accelerates the return of blood glucose levels in rats to basal level in the glucose tolerance test. The dose of Pep (6-13) was 200 μg/Kg body weight. Similarly, various forms of the peptide according to the invention in its A imide form potentiate the hypoglycaemic effect of insulin on "untrained" male and female mice. The effect on trained male mice trained to feed on schedule as described above was less striking, but still significant in some cases. These results are illustrated in Figures 12 and 13. As well as showing activity in the in vitro assays described above, peptides according to the invention showed hypoglycaemic activity in in vitro bioassays. Table 3 shows the effect of Pep (6-13) on glycogen deposition by isolated rat hemidiaphragms. There was not effect on the basal level in the absence of added insulin. At low or high insulin concentrations (10 2 and 104 μU/ml) respectively, Pep (6-13) A imide significantly increased glycogen deposition. The ^ form and the ring opened, hydrolysed A imide form were both inactive. TABLE 3 imide hyd = hydrolysed β imide form i.e. inactive ring opened form. The it imide form of the peptide also stimulated metabolism of glucose by isolated rat adipocytes, both in the presence and in the absence of added insulin. This is shown in Figure 14. The degree of stimulation depended on insulin dose. The dose of peptide was 10 μg per 3 ml culture. The effect of the A imide form peptides on glucose metabolism is further shown by their ability to stimulate glucose uptake and glycogen synthesis by cultured muscle cells and isolated rat hemidiaphragms. The results are presented in Tables 4 and 5. TABLE 4 In Vitro Effect of β-Imido Linked L-Aspartyl-L-Asparaginyl Peptides On Glucose Uptake In Rat Hemidiaphragms. TABLE Results of Cultured Cell Assays for the Stimulation of Glucose " Uptake " by Synthetic Peptides (.tyig/ml) Results of Isolated Muscle Assays,for the Stimulation of Glycogen Synthesis by Synthetic Peptides (lOOyg/ml) Peptide Rate of Increase from Basal (junto 1 glycogen/,-?: tissue/h ) P051 Leu-Ser-Arg- Leu-Phe-Λsp-Asn- la 0.74 P076 Leu-Ser-Λrg- Leu-Phe-Asp-Gln-Ala 0.30 P078 Leu-Ser-Arg- Leu-His-Asp-Asn-Λla 0.1 1 P087 Leu-Ser-Arg- •Ile-Phe-Asp-Asn-Ala 0.36 P0 0 Leu-Ser-Lys- • eu-Phe-Asp-Asn-Λla 0.16 P197 Cys-Tyr-Leu- •S e r-Arg-Leu-Phe-Λsp~Asn-As n 0.17 P202 Cys-Tyr-leu- •Ser-Arg-Leu-Phe-Asp-Asn-Ala 0.44 Of the peptides tested, Pep (1-15), (6-13), and (8-13) showed a highly significant stimulation; Pep (7-13) was on the borderline of significance, while Pep (9-13) and Pep (10-13) showed no effect. The results show that peptides of general formula I induce in a dose dependent manner a significant lowering of blood glucose levels at or below the level of lOμM/Kg body weight with the intravenous insulin tolerance test, and enhance in a dose dependent manner glucose utilisation via activation of glycogen synthase, incorporation of glucose into glycogen and oxidation of glucose via pyruvate to carbon dioxide. A dose of as little as 0.05 uM/Kg body weight was effective with some peptides. APPLICATIONS OF THE INVENTION The β -imido-L-aspartyl-L-asparaginyl peptides of general formula I induce (a) dose-dependent lowering of blood glucose and a potentiation of glycogen synthesis in vivo (b) dose-dependent oxidation of glucose and the activation of glycogen synthase in vitro. This suggests that the fi -imido- -aspartyl- L-asparaginyl peptide analogues are potent hypoglycaemic agents, and may be useful as therapeutic agents for the control of glucose metabolism, since normalisation of blood glucose levels in insulin-independent diabetes mellitus (Type II) mammals with glucose intolerance and insulin resistance is achieved by control of carbohydrate metabolism via glucose utilisation, and since glucose utilisation can be achieved via two pathways, namely oxidation of glucose or incorporation of glucose into glycogen. Since potentiation of insulin action results in an decreased requirement for endogenous insulin, the use of the fl -imido-L-aspartyl—L-asparaginyl peptide analogues may permit control of hyperinsulinaemia in insulin-independent Type II diabetes in humans or streptozotocin-treated animals. (f) By analogy, the use of other synthetic peptide analogues containing the ^--imido-L-aspartyl-L-asparaginyl moiety of general formula I is likely to exhibit either agonistic or antagonistic properties in vitro or in vivo related to glucose utilisation and so provide methods for control of hypoglycaemia or hyperglycaemia, depending on their respective agonistic or antagonistic properties in control of glucose metabolism. It will be clearly understood that the invention in its general aspects is not limited to the specific details referred to hereinabove. The following words used hereinabove are trade marks: BIOGEL, NUJOL.

Claims: -- -- CLAIMS: 1. A compound of general formula I: O R 10 — R n — Ro — R-7 — R-r e - R 4 NH- N CHC0R-, - R - R, O wherein X is hydrogen, -CHC0NH 2 or -CH 2 CK 2 C0NH 2 ; each of R,, R 2 and R, is a L- oc.-amino acid, a d -amino acid or an £-amino acid; R. " is an L or D «-amino acid, a £-amino acid or an £—amino acid; and each of R,- to R, , is hydrogen or an L or D <^-amino acid, a ώ -amino acid or an < £-amino acid, or a pharmaceutically acceptable salt thereof. 2. A compound according to Claim 1 in which R. is a L or D oC-amino acid, a S-amino acid or an -amino acid. 3. A compound according to Claim 1 or Claim 2 in which each of R-, , R 2 and R- is selected from the group consisting of alanine, glycine and phenylalanine. 4. A compound according to any one of Claims 1 to 3, in which R. is a hydrophobic amino acid. 5. A compound according to Claim 4 in which R. is selected from the group consisting of L-phenylalanine, L-tyrosine, L-tryptophan and L-histidine. 6. A compound according to any one of the preceding claims, in which 5 is a hydrophobic amino acid. 7. A compound according to Claim 6 in which R.. is selected from the group consisting of L- or D- leucine, isoleucine and histidine. 8. A compound according to any one of the preceding claims, in which g is a basic amino acid. 9. A compound according to Claim 9, in which R fi is selected from the group consisting of L- or D- arginine, lysine, and histidine. 10. A compound according to any one of the preceding claims, in which R-, is a substantially hydrophobic amino acid. 11. A compound according to Claim 10 in which R-, is selected from the group consisting of L- or D- serine, leucine and isoleucine. 12. A compound according to any one of the preceding claims, in which R g is selected from the group consisting of L- or D- leucine, phenylalanine, proline or isoleucine, an & -amino acid such as 6-aminohexanoic acid, and 4-aminocyclohexane-l-carboxylic acid. 13. A compound according to any one of the preceding claims, in which each of R g and R 10 is a substantially hydrophobic amino acid. 14. A compound according to Claim 13, in which each of g and R-, Q is selected from the group consisting of L- or D- leucine, phenylalanine, proline, isoleucine or tyrosine.. 15. A compound according to any one of the preceding claims, in which one of R g , R g or R-, Q is coupled to R, via a bifunctional agent. 16. A compound according to Claim 15, in which the bifunctional agent is 6-aminohexanoic acid. 17. A compound according to any one of the preceding claims, in which one or more of the amino acids has side chain substituents. 18. A compound according to any one of the preceding claims, in which R, , R 2 , R„, R g , or R 10 is absent from the peptide, allowing coupling between C-terminal and N-terminal amino acids via a bifunctional agent such as 6-aminohexanoic acid. 19. A pharmacologically active analogue or derivative of a compound according to any one of the preceding claims, in which the aspartimide ring is replaced by a group selected from o-aminobenzoic acid, m-aminobenzoic acid, p-aminobenzoic acid, o -lactams and ό-lactams. 20. A pharmaceutical composition comprising a compound according to any one of the preceding claims, together with a pharmaceutically acceptable carrier, diluent or excipient. 21. A method of solid phase synthesis of a peptide of general formula I, in which the peptide is bound to a resin, comprising the steps of: (a) protecting the od-amino functionality of added amino acids using t-BOC or f-MOC, (b) treating the resin-bound protected © / straight chain L-aspartyl-L-asparaginyl peptide with a base to produce the stable p-imido form of the peptide, and (c) recovering the peptide. 22. A method according to Claim 21, in which the base is a dialkylamine or a trialkylamine. 23. A method according to Claim 22, in which the base is piperidine or triethylamine. 24. A method of lowering the level of blood glucose in a mammal in need of such treatment, comprising administering to that animal a hypoglycaemically effective dose of a compound of general formula I, optionally together with or in conjunction with a second hypoglycaemic agent.

Inventors: HEARN MILTON THOMAS WILLIAM, NG FRANK MAN-WOON, O'DONOGHUE MICHAEL FRANCIS, RAE IAN DAVID, ROBSON VICTORIA MARIE JANE, HEARN, MILTON, THOMAS, WILLIAM, NG, FRANK, MAN-WOON, O'DONOGHUE, MICHAEL, FRANCIS, RAE, IAN, DAVID, ROBSON, VICTORIA, MARIE, JANE
Applicants: AUSTRALASIAN DRUG DEV LTD, UNIV MONASH, AUSTRALASIAN DRUG DEVELOPMENT LIMITED, MONASH UNIVERSITY
Application year: 1988
Main classification: C07K7
IPC codes: A61K38, C07K5, C07K14