Here we describe the design and application of OSu-FMS-MAL-S-(CH(2))(15)-COOH, an agent that associates with albumin while linked to a peptide or a protein with sufficient affinity (Ka=2 to 2.6 x 10(5)M(-1)) to protract the action of short- lived peptides and proteins in vivo. Under physiological conditions this probe undergoes spontaneous hydrolysis with the concomitant reactivation of inactive conjugates. Intravenously administered (125)I-labeled-Insulin-FMS-MAL-S-(CH(2))(15)-COOH to rats shows half-life of 17+/-2h, exceeding 5.2 times that obtained with intravenously administered (125)I-labeled Insulin. In mice this derivative facilitates glucose-lowering effect over a period of 24h, yielding AUC five times greater than that obtained by a similar dose of insulin-detemir. Similarly, subcutaneous administration of Exendin-4-FMS-MAL-S-(CH(2))(15)-COOH into mice facilitated prolonged and stable reduction in glucose level, yielding a t(1/2) value of 28+/-2h, exceeding the effect of exendin-4 4.7 folds. The inactive derivative gentamicin-FMS-MAL-S-(CH(2))(15)-COOH regained its full antibacterial potency upon incubation at physiological conditions yielding a t(1/2) value of 7.1+/-0.2h. In conclusion, the albumin-binding probe we introduced enables to prolong the action of any amino containing molecule in vivo, without the drawback of inactivation that often occurs upon such derivatization.

We attempted to engineer a novel long-acting insulin based on the following properties: (i) action as a prodrug to preclude supraphysiological concentrations shortly after injection; (ii) maintenance of low-circulating level of biologically active insulin for prolonged period; and (iii) high solubility in aqueous solution. A spontaneously hydrolyzable prodrug was thus designed and prepared by conjugating insulin through its amino side chains to a 40kDa polyethylene glycol containing sulfhydryl moiety (PEG(40)-SH), employing recently developed hetero-bifunctional spacer 9-hydroxymethyl-7(amino-3-maleimidopropionate)-fluorene-N-hydroxysucinimide (MAL-Fmoc-0Su). A conjugate trapped in the circulatory system and capable of releasing insulin by spontaneous chemical hydrolysis has been created. PEG(40)-Fmoc-insulin is a water-soluble, reactivatable prodrug with low biological activity. Upon incubation at physiological conditions, the covalently linked insulin undergoes spontaneous hydrolysis at a slow rate and in a linear fashion, releasing the nonmodified immunologically and biologically active insulin with a t(1/2) value of 30h. A single subcutaneous administration of PEG(40)-Fmoc-insulin to healthy and diabetic rodents facilitates prolonged glucose-lowering effects 4- to 7-fold greater than similar doses of the native hormone. The beneficial pharmacological features endowed by PEGylation are thus preserved. In contrast, nonreversible, "conventional" pegylation of insulin led to inactivation of the hormone.

Most peptide and protein drugs are short-lived species in vivo with a circulatory half-life of several minutes. This is particularly valid for non-glycosylated proteins with a molecular mass of less than 50 kDa. Since peptide/protein drugs are not absorbed orally, prolonged maintenance of therapeutically active drugs in the circulatory system is of primary clinical importance. Another major obstacle of injected polypeptide drugs is the elevated concentration of 100–1000 times above the therapeutical level that may be present in the circulatory system shortly after administration. Such overdosing may lead to undesirable side effects such as over-stimulation or down-regulation of receptor sites.

Annals of the New York Academy of SciencesVolume 805, Issue 1 p. 159-169 Development of VIP Agonists and Antagonists with Tissue and Receptor Specificity: Effects on Behavioral Maturation, Sexual Function, and the Biologic Clocka I. GOZES, Corresponding Author I. GOZES Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, Israel Tel: 972-3-640-7240; fax: 972-3-640-8541; e-mail: igozes@post.tau.ac.il.Search for more papers by this authorG. LILLING, G. LILLING Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. DAVIDSON, A. DAVIDSON Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. BARDEA, A. BARDEA Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. RESHEF, A. RESHEF Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorR. GLAZER, R. GLAZER Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorR. ZAMOSTIANO, R. ZAMOSTIANO Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorO. ASHUR-FABIAN, O. ASHUR-FABIAN Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. TICHER, A. TICHER Department of Human Genetics Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorI. E. ASHKENAZI, I. E. ASHKENAZI Department of Human Genetics Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorT. W. MOODY, T. W. MOODY Biomarkers and Prevention Research Branch NCI National Institutes of Health Rockville, Maryland 20850Search for more papers by this authorS. RUBINRAUT, S. RUBINRAUT Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorM. FRIDKIN, M. FRIDKIN Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorD. E. BRENNEMAN, D. E. BRENNEMAN Section on Developmental and Molecular Pharmacology Laboratory of Developmental Neurobiology National Institute of Child Health and Human Development National Institutes of Health Bethesda, Maryland 20842Search for more papers by this author I. GOZES, Corresponding Author I. GOZES Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, Israel Tel: 972-3-640-7240; fax: 972-3-640-8541; e-mail: igozes@post.tau.ac.il.Search for more papers by this authorG. LILLING, G. LILLING Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. DAVIDSON, A. DAVIDSON Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. BARDEA, A. BARDEA Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. RESHEF, A. RESHEF Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorR. GLAZER, R. GLAZER Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorR. ZAMOSTIANO, R. ZAMOSTIANO Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorO. ASHUR-FABIAN, O. ASHUR-FABIAN Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. TICHER, A. TICHER Department of Human Genetics Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorI. E. ASHKENAZI, I. E. ASHKENAZI Department of Human Genetics Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorT. W. MOODY, T. W. MOODY Biomarkers and Prevention Research Branch NCI National Institutes of Health Rockville, Maryland 20850Search for more papers by this authorS. RUBINRAUT, S. RUBINRAUT Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorM. FRIDKIN, M. FRIDKIN Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorD. E. BRENNEMAN, D. E. BRENNEMAN Section on Developmental and Molecular Pharmacology Laboratory of Developmental Neurobiology National Institute of Child Health and Human Development National Institutes of Health Bethesda, Maryland 20842Search for more papers by this author First published: December 1996 https://doi.org/10.1111/j.1749-6632.1996.tb17481.xCitations: 4 a This work was supported in part by Fujimoto Pharmaceutical Corporation, and in part by the Israel-U.S. Binational Science Foundation. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume805, Issue1VIP, PACAP, Glucagon, and Related PeptidesDecember 1996Pages 159-169 RelatedInformation

The covalent linkage of peptides or protein drugs to human serum albumin (HSA) greatly prolongs their lifetime in vivo but is pharmacologically irrelevant when it irreversibly inactivates them. We retain drug bioactivity by synthesizing a heterobifunctional reagent (MAL-Fmoc-OSu, 9-hydroxymethyl-2-(amino-3-maleimidopropionate)-fluorene-N-hydroxysuccinimide) that generates HSA-Fmoc-insulin on covalent conjugation to the amino group of insulin and the Cys-34 side chain of HSA. HSA-Fmoc-insulin is water-soluble and, upon incubation in aqueous buffers reflecting normal human serum conditions, slowly, spontaneously, and homogeneously hydrolyzes to release unmodified insulin with a t 1/2 of 25 +/- 2 h. A single subcutaneous or intraperitoneal administration of HSA-Fmoc-insulin to diabetic rodents lowers circulating glucose levels for about 4 times longer than an equipotent dose of Zn2+-free insulin. Following subcutaneous administration, onset of the glucose-lowering effect is delayed 0.5-1 h and persists for 12 h. Thus, we present a prototype insulin formulation possessing three desirable parameters: high aqueous solubility, delayed action following subcutaneous administration, and prolonged therapeutic effect.

Alzheimer's disease (AD) is characterized by death of selected brain cells that normally supply messenger molecules, forming short-term memory. The focus of the current work is the discovery and characterization of very short, readily available proteins (termed peptides) that provide protection to nerve cells at very low concentrations (among the lowest described to date).A major brain natural-protective short protein is vasoactive intestinal peptide (VIP) that affords neuronal defense through activation of proteins derived from glial cells (the brain support cells), such as activity-dependent neurotrophic factor (ADNF) and activity-dependent neuroprotective protein (ADNP). Drug design identified small and modified peptide fragments derived from VIP, ADNF, and ADNP, which can protect against damage in cell and animal models. The specific aim of the study is to optimize and choose the lead compounds. Tests include: simplified tissue-culture systems, brain-penetration assessments, and effects on short-term memory, in models relevant to AD. Selected lead compounds are further tested for mechanisms of protection, as well as potential side effects, and put forth for future clinical evaluations.Key findings include: (1) Lead compounds have been chosen: (a) the ADNP-derived peptide, NAP, (b) the fat-modified VIP, SNV, (c) the shortened VIP-derivative, stearyl-KKYL-NH2, and (d) the ADNF-related peptide, ADNF-9; (2) formulations for nasal spray administration have been optimized; (3) studies on mechanisms of protection identified selective gene activation and defense against inflammation. Advanced geneticengineering techniques are now utilized for the identification of novel interacting molecules, with NAP, the first lead planned for clinical trials, as the target.

Vasoactive intestinal peptide (VIP) is a recognized growth factor affecting many cell types. We have previously developed a series of lipophilic VIP analogues containing an N-terminal covalently attached stearyl moiety. The current studies identified stearyl-Nle(17)-VIP and stearyl-Nle(17)-neurotensin(6-11)VIP(7-28), acting at microM concentrations, as cytotoxic to human keratinocytes. The core C-terminal active VIP-derived peptide, stearyl-Lys-Lys-Tyr-Leu-NH(2) (St-KKYL-NH(2)), was identified as being responsible for the observed cytotoxicity. Cytotoxicity coincided with marked reduction in intracellular cyclic GMP and was abolished by co-treatment with the endonuclease inhibitor, aurine-tricarboxylic acid, suggesting apoptotic mechanisms. Stearyl-VIP derivatives thus offer lead compounds for future drug development against hyperproliferative skin conditions.

The understanding of the molecular mechanisms leading to peptide action entails the identification of a core active site. The major 28-aa neuropeptide, vasoactive intestinal peptide (VIP), provides neuroprotection. A lipophilic derivative with a stearyl moiety at the N-terminal and norleucine residue replacing the Met-17 was 100-fold more potent than VIP in promoting neuronal survival, acting at femtomolar-picomolar concentration. To identify the active site in VIP, over 50 related fragments containing an N-terminal stearic acid attachment and an amidated C terminus were designed, synthesized, and tested for neuroprotective properties. Stearyl-Lys-Lys-Tyr-Leu-NH2 (derived from the C terminus of VIP and the related peptide, pituitary adenylate cyclase activating peptide) captured the neurotrophic effects offered by the entire 28-aa parent lipophilic derivative and protected against beta-amyloid toxicity in vitro. Furthermore, the 4-aa lipophilic peptide recognized VIP-binding sites and enhanced choline acetyltransferase activity as well as cognitive functions in Alzheimer's disease-related in vivo models. Biodistribution studies following intranasal administration of radiolabeled peptide demonstrated intact peptide in the brain 30 min after administration. Thus, lipophilic peptide fragments offer bioavailability and stability, providing lead compounds for drug design against neurodegenerative diseases.

Abstract: Pituitary stimulating adenylate cyclase (PACAP) is a major regulatory peptide with two active molecular forms: PACAP‐27 and PACAP‐38. Both molecular forms promote neuronal survival and protect against neurotoxicity. Based on our previous hybrid peptide strategy in designing vasoactive intestinal peptide (VIP) antagonists, novel PACAP analogues were synthesized (neurotensin6‐11 PACAP7‐27 and neurotensin6‐11 PACAP7‐38). In addition to the hybrid modification, the methionine in position 17 was replaced by norleucine (Nle). Treatment of rat cerebral cortical cultures for five days with the putative PACAP antagonists (1 nM) resulted in a 35‐45% reduction in neuronal cell counts as compared to controls. Neuronal cell death was already obtained at picomolar concentrations for the neurotensin6‐11PACAP7‐27 antagonist with 70% death at 10−8 M. Co‐administration of the PACAP hybrid analogue with picomolar amounts of PACAP‐27 or Nle17‐PACAP‐27 attenuated the reduction in neuronal cell counts. While the protective effects of both analogues exhibited a peak at 1 pM concentrations, the Nle‐containing agonist displayed a broader range of active concentrations (10−12 M‐10−9 M)The putative PACAP antagonist also inhibited sperm motility (golden hamster) in a dose‐dependent manner as assessed in vitro. Complete inhibition was observed at 10 μM, suggesting a role for PACAP in sperm motility and sexual function. Thus, previous findings of a large number of PACAP and PACAP receptors in the nervous system and the reproductive system are now correlated with a function in neuronal survival and sperm motility. The structure‐activity studies suggest that the methionine in position 17 and the first six amino acids are important in the determination of PACAP activity, knowledge that may facilitate PACAP‐based drug design.

Activity-dependent neurotrophic factor (ADNF) is a glia-derived protein that is neuroprotective at femtomolar concentrations. A 14-amino acid peptide of ADNF (ADNF-14) has been reported that protects cultured neurons from multiple neurotoxins. Structure-activity relationships of peptides related to ADNF-14 now have been determined. A 9-amino acid core peptide (ADNF-9) has been identified that has greater potency and a broader effective concentration range (10(-16) to 10(-13) M) than ADNF or ADNF-14 in preventing cell death associated with tetrodotoxin treatment of cerebral cortical cultures. Deletions or conservative amino acid substitutions to ADNF-9 resulted in reduced potency, narrower effective concentration range and/or decreased efficacy. Removal of the N-terminal serine or the COOH-terminal isoleucine-proline-alanine from ADNF-9 produced a significant reduction in survival-promoting activity. Comparative studies of ADNF-9 action in mixed (glia plus neurons) vs. glia-depleted neuronal cultures indicated that ADNF-9 can act directly on neurons, although the potency of the peptide was 10,000-fold greater in mixed cultures. Kinetic studies showed that exposure to ADNF-9 for only 2 hr was sufficient to produce a 4-day protection against the cell-killing action of tetrodotoxin. Treatment with bafilomycin A1 (an inhibitor of receptor-mediated endocytosis) for 2 hr prevented the ADNF- and ADNF-9-mediated neuroprotection. ADNF-9, like ADNF-14, was neuroprotective against N-methyl-D-aspartate and the beta-amyloid peptide (amino acids 25-35), and had a much broader range of effective concentrations than ADNF-14. These studies identify ADNF-9 as an attractive lead compound for the development of therapeutic agents against neurodegenerative diseases.

Neuropeptides generally exhibit multiple roles in the maintenance of homeostasis. Classically, neuropeptides were found to exert neurohormonal and neurotransmitter (neuromodulator) effects in the central and peripheral nervous system. It is becoming increasingly apparent that neuropeptides also act as regulators of cell division, differentiation, and survival. In the past we reviewed neuropeptides as growth factors.'Z2 Recent studies indicate that neuronal cells, in response to axotomy, increase expression of neuropeptides that are associated with the promotion of survival and regeneration (see ref. 3 for review). Examples include corticotropin-releasing factor, dynorphin, calcitonin gene-related peptide, cholecystokinin, galanin, vasoactive intestinal peptide (VIP), neuropeptide Y, and others. At the same time, down-regulation of neurotransmitters and substances related to the secretion of neurotransmitters may occur. Similarly, neurotrophins of the family of nerve growth factors (NGF) increase the expression of specific neuropeptides, for example, brain-derived neurotrophic factor enhanced neuropeptide Y and somatostatin levels in cortical neurons, whereas NGF did not.4 In the periph-

Stearyl-Nle17-VIP (SNV) is a novel agonist of vasoactive intestinal peptide (VIP) exhibiting a 100-fold greater potency than the parent molecule and specificity for a receptor associated with neuronal survival. Here, mice deficient in apolipoprotein E (ApoE), a molecule associated with the etiology of Alzheimer's disease, served as a model to investigate the developmental and protective effects of SNV. In comparison to control animals, the deficient mice exhibited (a) reduced amounts of VIP messenger RNA; (b) decreased cholinergic activity (c) significant retardation in the acquisition of developmental milestones: forelimb placing behavior and cliff avoidance behavior; and (d) learning and memory impairments. Daily injections of SNV to ApoE-deficient newborn pups resulted in increased cholinergic activity and marked improvements in the time of acquisition of behavioral milestones, with peptide-treated animals developing as fast as control animals and exhibiting improved cognitive functions after cessation of peptide treatment. Specificity was demonstrated in that treatment with a related peptide (PACAP), pituitary adenylate cyclase-activating peptide, produced only limited amelioration. As certain genotypes of ApoE increase the probability of Alzheimer's disease, early counseling and preventive treatments may now offer an important route for therapeutics design.

The effects of pituitary adenylate cyclase activating polypeptide (PACAP) hybrid, a synthetic antagonist, was investigated on NIH/3T3 cells containing PACAP receptor (R) splice variants (SVs). PACAPhybrid inhibited 125I-PACAP-27 binding to NIH/3T3 cells stably expressing PACAP-R basic, SV-1, SV-2 or SV-3 with an IC50 of 1000 nM. PACAPhybrid antagonized the ability of PACAP-27 to elevate cAMP regardless of the PACAP-R SV used. PACAP was more efficacious at increasing cytosolic Ca2+ in NIH/3T3 cells containing PACAP-R SV-2 than PACAP-R basic, SV-1 or SV-3. PACAPhybrid antagonized the increase in cytosolic Ca2+ caused by PACAP-27 regardless of the PACAP-R SV used. PACAP was more potent at elevating c-fos mRNA using NIH/3T3 cells transfected with PACAP-R SV-2 than PACAP-R basic, SV-1 or SV-3. PACAPhybrid antagonized the increase in c-fos mRNA caused by PACAP-27. These data suggest that PACAPhybrid is a useful PACAP receptor antagonist for PACAP-R SVs.

Annals of the New York Academy of SciencesVolume 814, Issue 1 p. 161-166 Neuropeptides and Neuronal Survival: Neuroprotective Strategy for Alzheimer's Diseasea I. GOZES, Corresponding Author I. GOZES Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, Israele Corresponding author.Search for more papers by this authorA. BARDEA, A. BARDEA Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorM. BECHAR, M. BECHAR Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorO. PEARL, O. PEARL Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. RESHEF, A. RESHEF Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorR. ZAMOSTIANO, R. ZAMOSTIANO Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. DAVIDSON, A. DAVIDSON Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorS. RUBINRAUT, S. RUBINRAUT Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorE. GILADI, E. GILADI Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorM. FRIDKIN, M. FRIDKIN Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorD. E. BRENNEMAN, D. E. BRENNEMAN Section on Developmental and Molecular Pharmacology Laboratory of Developmental Neurobiology National Institute of Child Health and Human Development National Institutes of Health Bethesda, Maryland 20892Search for more papers by this author I. GOZES, Corresponding Author I. GOZES Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, Israele Corresponding author.Search for more papers by this authorA. BARDEA, A. BARDEA Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorM. BECHAR, M. BECHAR Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorO. PEARL, O. PEARL Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. RESHEF, A. RESHEF Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorR. ZAMOSTIANO, R. ZAMOSTIANO Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorA. DAVIDSON, A. DAVIDSON Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorS. RUBINRAUT, S. RUBINRAUT Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorE. GILADI, E. GILADI Department of Clinical Biochemistry Sackler School of Medicine Tel Aviv University Tel Aviv 69978, IsraelSearch for more papers by this authorM. FRIDKIN, M. FRIDKIN Department of Organic Chemistry Weizmann Institute of Science Rehovot 76100, IsraelSearch for more papers by this authorD. E. BRENNEMAN, D. E. BRENNEMAN Section on Developmental and Molecular Pharmacology Laboratory of Developmental Neurobiology National Institute of Child Health and Human Development National Institutes of Health Bethesda, Maryland 20892Search for more papers by this author First published: 17 December 2006 https://doi.org/10.1111/j.1749-6632.1997.tb46154.xCitations: 15 a This research was supported in part by the Fujimoto Corporation and the U.S.-Israel Binational Science Foundation. ‡ This paper was written while I.G. was a scholar-in-residence at the Fogarty International Center for Advanced Study in the Health Sciences, National Institutes of Health, Bethesda, Maryland. Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume814, Issue1Neuropeptides in Development and AgingApril 1997Pages 161-166 RelatedInformation

Neurodegenerative diseases, in which neuronal cell disintegrate, bring about deteriorations in cognitive functions as is evidenced in millions of Alzheimer patients. A major neuropeptide, vasoactive intestinal peptide (VIP), has been shown to be neuroprotective and to play an important role in the acquisition of learning and memory. A potent lipophilic analogue to VIP now has been synthesized, [stearyl-norleucine17]VIP ([St-Nle17]VIP), that exhibited neuroprotection in model systems related to Alzheimer disease. The beta-amyloid peptide is a major component of the cerebral amyloid plaque in Alzheimer disease and has been shown to be neurotoxic. We have found a 70% loss in the number of neurons in rat cerebral cortical cultures treated with the beta-amyloid peptide (amino acids 25-35) in comparison to controls. This cell death was completely prevented by cotreatment with 0.1 pM [St-Nle17]VIP. Furthermore, characteristic deficiencies in Alzheimer disease result from death of cholinergic neurons. Rats treated with a cholinergic blocker (ethylcholine aziridium) have been used as a model for cholinergic deficits. St-Nle-VIP injected intracerebroventricularly or delivered intranasally prevented impairments in spatial learning and memory associated with cholinergic blockade. These studies suggest both an unusual therapeutic strategy for treatment of Alzheimer deficiencies and a means for noninvasive peptide administration to the brain.

To distinguish vasoactive intestinal peptide (VIP) receptors in the brain-mediating neurotransmission and neurotrophism, potent VIP analogues were designed. Using a single amino acid substitution and the addition of a fatty acyl moiety, an analogue was devised that exhibited both a 100-fold greater potency than VIP and specificity for a VIP receptor associated with neuronal survival. This VIP agonist increased neuronal survival via a cAMP-independent mechanism. Identical chemical modification of a prototype VIP antagonist (Met-Hybrid, Neurotensin6-11-VIP7-28) also resulted in a 100-fold greater potency in blocking VIP-mediated increases in neuronal survival. Blockade of circadian activity rhythms was limited to VIP antagonists that could inhibit VIP-mediated increases in cAMP. These lipophilic peptides provide novel tools in receptor discrimination and drug design.