Biomedical subjects
Publications and source records attributed to W Haq.
Solid phase syntheses of ethyl 6-carboxyquinol-4(1H-)-one-3-carboxylate (5) and N-substituted 6-carboxyquinol-4(1H)-one-3-carboxamides 7a-d have been described. Antifilarial in vitro activities of 5,7a-d against Brugia malayi have also been delineated.
An efficient solid phase synthesis of pyrimido[4,5-d]pyrimidine derivatives is described. Reaction of polymer-bound pyrimidine 1 with urea or thiourea followed by cleavage from the support provided 4-aminopyrimido[4,5-d]pyrimidines 4 and 5 while treatment of 6 with phenyl isocyanate or phenyl isothiocyanate followed by cleavage from resin afforded 3-phenylpyrimido[4,5-d]pyrimidines 9 and 10.
Thymopentin and its analogs have been synthesized by the solution phase method of peptide synthesis and evaluated for their prophylactic efficacy against L. donovani infection in hamsters. Thymopentin and some of the analogs were found to stimulate nonspecific resistance of the host against Leishmania donovani infection in hamsters.
A novel general synthesis of substituted pyrimidine 3 has been carried out on solid support. The C-atoms carring the cyano, amino, carboxamido, as well as anchoring site have exploited to generate libraries of compounds 6-8, 10, 13, 15, 17, 19, 21, 23, 25 and 27. A novel strategy to cleave the resin to resin-site unsubstituted system has been developed and it provides 5,6-disubstituted pyrimidines 6-8. In addition, synthesis of 2,5,6-trisubstituted pyrimidines of prototype 10 were carried out by nucleophilic displacement of the anchor by various amines. Further investigations were directed toward the solid phase synthesis of pyrimido[4,5-d]pyrimidines 12, 16, 20 and 24 in which C-atoms carring the oxo, thio, amino, anchoring site as well as NH could be introduced as center of diversity to generate libraries of compounds for potential use. 4-Aminopyrimido[4,5-d]pyrimidines 13 and 17 were obtained from fusion of 3a with urea or thiourea followed by cleavage of support while 3-phenylpyrimido[4,5-d]pyrimidines 21 and 27 were synthesized from cyclisation of 4 with phenyl isocyanate or isothiocyanate followed by release of resin. 7-substituted pyrimido[4,5-d]pyrimidines 15, 19, 23 and 27 were obtained by oxidation of 12, 16, 20 and 24 followed by aminolytic cleavage of support.
Modifications of 4,4' residues of Biphalin have resulted in greater binding selectivity and biological potency for the mu opioid receptor. A higher partition coefficient across the phospholipid bilayer membrane has been achieved by using a beta-branched unusual amino acids.
Human beta-casein fragment (54-59) having the amino acid sequence Val-Glu-Pro-Ile-Pro-Tyr, has been shown potent immunostimulant activity. Several analogs of this hexapeptide have been synthesized with modification in the N-terminal region and tested for their immunomodulatory activity. Interestingly, two hexapeptides have shown significant immunosuppressant activity.
Splenopentin (SP-5, Arg-Lys-Glu-Val-Tyr) and thymopentin (TP-5, Arg-Lys-Asp-Val-Tyr) are synthetic immunomodulating peptides corresponding to the region 32-34 of a splenic product called splenin (SP) and the thymic hormone thymopoietin (TP), respectively. TP was originally isolated as a 5-kDa (49-amino acids) protein from bovine thymus while studying effects of the thymic extracts on neuromuscular transmission and was subsequently observed to affect T cell differentiation and function. TP I and II are two closely related polypeptides isolated from bovine thymus. A radioimmunoassay for TP revealed a crossreaction with a product found in spleen and lymph node. This product, named splenin, differs from TP only in position 34, aspartic acid for bovine TP and glutamic acid for bovine splenin and it was called TP III as well. Synthetic pentapeptides (TP-5) and (SP-5), reproduce the biological activities of TP and SP, respectively. It is now evident that various forms of TPs were created by proteolytic cleavage of larger proteins during isolation. cDNA clones have been isolated for three alternatively spliced mRNAs that encodes three distinct human T cell TPs. The immunomodulatory properties of TP, SP, TP-5, SP-5 and some of their synthetic analogs reported in the literature have been briefly reviewed.
Seven new tetrapeptides analogous to (1-4) sequence of dermorphin were synthesized and evaluated for their opioid activity. The peptides were synthesized by the solution phase method. Their opioid activity revealed that peptides II and V were the most potent in the analgesia test as well as in the peripheral assays. Peptide II was most active in the guinea pig ileum assay, whereas peptide VI was 2763 times more selective for mu-receptors.
The synthetic nonapeptide Val-Gln-Gly-Glu-Glu-Ser-Asn-Asp-Lys corresponding to the amino acid sequence 163-171 of human interleukin-1beta (IL-1beta) has been reported to retain considerable immunostimulatory activity of the native protein without the induction of the inflammatory or pyrogenic responses. Two lipophilic derivatives of this nonapeptide, one having a lauroyl residue (1) and the other having a palmitoyl residue (2) at the N-terminus of the peptide, and a more stable analog carrying D-Val residue at position 1 of the peptide (3) were synthesized with a view to find out if these structural modifications had a favorable effect on in vitro mouse thymocyte proliferation and IL-1 dependent inhibition of A375 cells. We have found that analogs (1) and (2) are active in both the tests like the parent nonapeptide. The lipophilic analog (2) is in fact, effective at a lower dose as compared to the parent nonapeptide in mouse thymocyte proliferation assay. Although the analog (3) has the ability to inhibit A375 cells, it does not stimulate mouse thymocyte proliferation in vitro. The IL-1beta fragment (163-171) and the analog (2) were further compared for their effects on pyrogenicity, blood glucose level, acute phase response and radioprotection. Unlike IL-1beta, its fragment (163-171) and the analog (2) do not induce pyrogenicity and any of the acute phase related changes such as the increase in C-reactive protein and hypoglycemia following their administration in Balb/c mice. We have found that 40% of animals treated with analog (2) survive more than 21 days after lethal irradiation as compared to 20% survivors in groups treated with recombinant IL-1beta or its nonapeptide fragment (163-171), under conditions when all the control animals died within 10 days. This study may help in designing small peptides which may be more effective and stable.
Synthetic peptide antigens representing the repeat sequences of malarial antigens showed poor immunogenicity and protection in clinical trials. In the present study, RESA, an asexual blood stage antigen, containing (EENVEHDA)2 and (DDEHVEEPTVA)2 sequences were chemically linked to a promiscous T-cell determinant (CS.T3) of the circumsporozoite protein of P. falciparum. The synthetic constructs either alone or coentrapped with immunoadjuvants (nor muramyl dipeptide/lauroyl tetrapeptide) were administered in liposomes to mice of varying genetic background and the immunogenicity of different formulations were compared under identical experimental conditions. The RESA peptide formulations containing the T-cell determinant and the adjuvants generated high titre and affinity antibodies in all the strains, as compared to peptide(s) alone. The booster immunization induced a strong anamnestic response in each group. Though the major IgG isotype is of IgG1 and IgG2b interestingly, formulations containing CS.T3 have a higher proportion of cytophilic IgG2b isotype. There was a significant fall in the levels of IgG2b isotype while IgG1 levels were maintained same in the third bleed (day 60, without booster immunization). The mixed peptide group preparation containing the adjuvant is found to be a better immunogen than that of respective peptides itself. The in vitro merozoite reinvasion inhibition assay showed 76-96% inhibition with the formulations containing RESA peptide(s)-CS.T3 and the adjuvant, while with peptides alone the inhibition was 50-56%. This study highlights the importance of an alternative approach for developing peptide based immunogen against malaria.
An up-to-date review is presented covering all the available information concerning the isolation, discovery, synthesis, conformation, receptor binding characteristics, pharmacological properties and SAR studies of dynorphin A and related peptides. The potential of dynorphin A and its analogs has yet to be fully realized.
Met-enkephalin (Tyr-Gly-Gly-Phe-Met) and its more stable analogs, Tyr-D-Ala-Gly-MePhe-Met-NHC3H gamma-iso (1) and Tyr-D-Ala-Gly-MePhe-Gly-NHC3H gamma-iso (2) significantly enhanced human T-cell proliferation in vitro after 5 days of incubation in the absence of mitogen. The activity was completely inhibited by naloxone, an opioid antagonist. These peptides significantly enhanced human active T-cell rosette (CD2R) also on in vitro treatment. Furthermore, these analogs stimulated interleukin-2 production by human peripheral blood mononuclear cells in vitro which was completely inhibited by naloxone. These observations suggest that human T-cells bear receptors for Met-enkephalin on their surface. Such findings may provide a link between the central nervous system and the immune system.
Splenopentin (SP-5), is a pentapeptide corresponding to the amino acid sequence 32-36 (Arg-Lys-Glu-Val-Tyr) of the splenic hormone splenin. Its synthetic analogs: Lys-Lys-Glu-Val-Tyr(1) and D-Lys-Lys-Glu-Val-Tyr (2) have been evaluated for active T-cell rosette (CD2R), total T-cell rosette (CD2), interleukin-2 (IL-2) stimulation and effect on antibody production. SP-5 as well as both the analogs stimulated CD2R. Analogs (1) and (2) were also found to stimulate IL-2 production. These observations suggest that in vitro human NK cell augmentation with analogs (1) and (2) reported earlier may be due to enhanced IL-2 production.
Structural analogs of the hexapeptide sequence 54-59 (A) human casein, reported to stimulate immune response, were synthesized and evaluated for immunostimulant activity. Hexapeptide 91/409 (C), 90/649 (D) and 91/361 (E) stimulated higher antibody titre and delayed type of hyper-sensitivity (DTH) response than the natural casein hexapeptide in BALB/c mice-sheep red blood cells (SRBC) and guinea pig-ovalbumin models. These peptides also induced higher stimulation of non-specific immune response as evidenced by increase in macrophage migration index (MMI), phagocytosis of (14C) lecuine labelled Escherchia coli, incorporation of (14C)-glucosamine in peritoneal macrophages and proliferative response of mouse thymocytes. Significant suppression on the course of Plasmodium berghei infection was also observed on day 4, in the animals treated with hexapeptidse C and D.
2-methyl-4a alpha-(3-hydroxyphenyl)-1,2,3,4,4a,5,12,12a alpha-octahydro-quinolino[2,3,30g]isoquinoline (TAN-67) is a nonpeptidic delta-opioid receptor agonist. This report describes its receptor binding affinity and agonist potency at human and mouse delta and mu-opioid receptors. The binding affinities of TAN-67 and the cyclic enkephalin analog, (D-Pen2, 4'-Cl-Phe4, D-Pen5]enkephalin (pCl-DPDPE) were measured by radioligand binding inhibition studies at mouse and human variants of the delta and mu-opioid receptor using [3H]Naltrindole and [3H]D-Phe-Cys-Tyr-D-Trp-Orn-Thr -Pen-Thr-NH2, respectively. TAN-67 showed high binding affinity (Ki = 0.647 nM) at the human delta-opioid receptor and high delta-opioid receptor binding selectivity ( > 1000-fold) relative to the human mu-opioid receptor. TAN-67 also showed high potency (EC50 = 1.72 nM) for the inhibition of forskolin-stimulated cAMP accumulation at human delta-opioid receptors expressed by intact Chinese hamster ovary cells but low potency (EC50 = 1520 nM) at human mu-opioid receptors expressed by intact B82 mouse fibroblast cells. The results show that TAN-67 has similar binding affinities, selectivity and potencies as pCl-DPDPE at human delta and mu-opioid receptors. These results combined with the nonpeptidic structure of TAN-67 suggest that this compound has therapeutic potential as a delta-opioid receptor agonist.
Several novel type of lipopeptides were synthesized and evaluated for their ability to stimulate non-specific resistance against Leishmania donovani infection. Peritoneal macrophages isolated from young male hamsters treated with muramyl dipeptide (MDP) and various synthetic lipopeptides (6 mg/kg i.p.) 7 days earlier, were cultured in vitro and challenged 24 h later with L. donovani promastigotes. One lipopeptide, Central Drug Research Institute (CDRI) compound 86/450, exhibited significantly higher immunostimulatory activity than MDP. Its prophylactic activity was further confirmed in hamsters by giving 2 split doses of 3 mg/kg of the compound spaced at 2 weeks, i.e. on day -7 and +7 of challenge with L. donovani amastigotes. The prophylactic effect lasted for 7 days following the last treatment with compound 86/450. The antileishmanial action of sodium stibogluconate (SAG) was also found to be enhanced by 16% in hamsters primed with compound 86/450.
Acute administration of cyclo (His-Pro) to rats cause a dose-dependent decrease in ethanol-induced hypothermia. Bromination of the imidazole moiety of histidine in cyclo (His-Pro) resulted in a significant increase in its potency to attenuate ethanol hypothermia. In contrast, benzylation of the imidazole moiety of histidine or the substitution of one or both of the amino acids in cyclo(His-Pro) led to a total loss of its thermomodulatory activity. In conclusion, it appears from these preliminary data that it may be possible to design analogs of CHP that may be effective antagonists for ethanol hypothermia.
The effects of Met-enkephalin (Met-Enk), a delta receptor binding opioid peptide, and its more stable synthetic analogs, Tyr-D-Ala-Gly-MePhe-Met-NHC3H7-iso (1), Tyr-D-Ala-Gly-MePhe-Gly-NHC3H7-iso (2) and Tyr-D-Ala-Gly-MePhe-Gly-NHCH2C6H5 (3), on human T-cell transformation and natural killer (NK) cell cytotoxicity have been evaluated. Analogs 1 and 2 have been found to be as potent as Met-Enk in stimulating T-cell transformation and augmenting NK cell cytotoxicity. Analog 3 had no effect on T-cell transformation and NK cell cytotoxicity. Proliferative response was measured by 3H-thymidine uptake after 5 days of incubation. The kinetics of the T-cell transformation response (peak 5th day) is similar to those for in vitro T-cell responses to specific antigens rather than via polyclonal activation.