A solid-phase synthesis of [lysine] vasopressin through a crystalline protected nonapeptide intermediate

Meienhofer, Johannes; Sano, Yoshimoto

doi:10.1021/ja01013a067

“…Total yields, based on the dipeptide content of the starting resin, were 20-25% in several syntheses. These yields were similar to those obtained in a standard solid-phase synthesis of lysine-vasopressin (33). The hormone, prepared from 6 by deprotection and oxidative cyclization, possessed units/mg of activity by a rat-pressor assay for different batches synthesized.…”

supporting

confidence: 76%

“…The pH was adjusted to 7.1 with 0.5 N NH40H and air was passed through the solution for 1 hr, after which the pH was adjusted to 4.8 with acetic acid. The solution was desalted on an Amberlite IRC-50 column (43), as described before (33,34), to give, after lyophilization, a colorless powder (54 mg, 87%) possessing a rat-pressor potency of 190 units/mg.…”

Section: Methodsmentioning

confidence: 99%

“…Purification of 46 mg by chromatography on an IRC-50 column in 0.5 M ammonium acetate buffer, pH 6.4 (36), as described earlier (33,34), gave lysine-vasopressin as a colorless powder (30 mg) with rat-pressor activity of 260-280 units/mg.…”

Section: Methodsmentioning

confidence: 99%

“…Deprotection by sodium in liquid ammonia, oxidative disulfide bond formation, desalting, and purification by ion-exchange chromatography (36) on Amberlite IRC-50 in 0.5 M ammonium acetate buffer, pH 6.4 (33,34) bonding to the solid phase, which would allow the removal of the completed peptide by milder treatments than the acidolytic reactions presently used, is strikingly apparent in the very recent solid-phase synthesis of human growth hormone (6), in which considerable activity was lost at this penultimate step of a total of 190 operational stages. The type of bonding used in this work allowed somewhat milder cleavage conditions compared with the standard Merrifield benzyl ester cleavage, but acidolytic treatment was still required.…”

mentioning

confidence: 99%

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Solid-Phase Synthesis with Attachment of Peptide to Resin through an Amino Acid Side Chain: [8-Lysine]-Vasopressin

Meienhofer

¹

,

Trzeciak

²

1971

Proc. Natl. Acad. Sci. U.S.A.

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It is proposed that the scope of solid-phase peptide synthesis could be considerably broadened by attaching peptides to the solid-phase through functional side-chain groups rather than through the commonly used a-carboxyl groups. Side-chain attachment offers the use of a large variety of chemical linkages to solid supports. Attachment through the e-amino group of the lysine residue to a polystyrene resin has been applied to a solid-phase synthesis of lysine-vasopressin. Na-tert-butyloxycarbonyl-L-lysyl-glycinamide was condensed with chloroformoxymethyl polystyrene-2% divinylbenzene resin. After removal of the Na-protecting tert-butyloxycarbonyl group, the peptide chain was elongated by standard Merrifield procedures to give Tos-Cys(Bzl)-Tyr-Phe-Glu-(NH2) -Asp(NH2) -Cys(Bzl) -Pro -Lys(Z -resin) -Gly-NH2. Cleavage from the resin with HBr in dioxane or trifluoroacetic acid gave a partially protected nonapeptide hydrobromide. For purification, it was converted into a fully protected peptide by treatment with benzyl p-nitrophenyl carbonate and crystallized. Deprotection by sodium in liquid ammonia, oxidative cyclization, IRC-50 desalting, and ion-exchange chromatography gave lysinevasopressin with high potency in a rat-pressor assay.The solid-phase method of peptide synthesis, developed by Merrifield (1, 2), makes possible the chemical syntheses of proteins. Synthetic achievements to date include syntheses of bovine insulin (3) in 1966, an analog of horse heart cytochrome c (4) and bovine pancreatic ribonuclease A (5) in 1969, and human growth hormone (6) in 1970. The advantages of the solid-phase method lie in (a) unprecedented speed, (b) simplicity of operation, (c) elimination of solubility problems, (d) minimal danger of racemization, and (e) possibility for automation. However, at its present state of development, the procedure suffers from several serious shortcomings. Incomplete peptide-bond formation (a), potentially occurring in each peptide-coupling step, results in so-called failure sequences and truncated sequences (7). The reagents (b) that must be used to remove the final product from the solid support often give incomplete cleavage and (or) partial degradation of the peptide. Analytical methods (c) sensitive enough to accurately monitor the process and to assess the purity of the products are not yet available.Some of the many efforts to improve the solid-phase method have focused on variations of the solid-phase (1, 8-16), or on facilitating formation of the commonly used benzyl-ester bond (17)(18)(19) between the starting C-terminal amino acid and the standard resin (chloromethyl polystyrene-2% divinylbenzene). In all of these investigations, the attachment to the solid-phase has been made through the a functional groups of amino acids or peptides (2). We wish to suggest that attachment to a solid-phase through functional groups in sidechains of amino acids could, (a) considerably broaden the chemical scope of such investigations, and (b) eliminate any danger of partial racemization of the am...

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“…Abbreviations of amino-acid derivatives and peptides are in accordance with the IUPAC-IUB Tentative Rules on Biochemical Nomenclature, Biochemistry, 5,2485,1445 (1966); 6,362 (1967). The amino acids (except glycine) had the L configuration unless otherwise stated.…”

mentioning

confidence: 99%

Regulation of Formation and Proposed Structure of the Factor Inhibiting the Release of Melanocyte-Stimulating Hormone

Celis

¹

,

Taleisnik

²

,

Walter

³

1971

Proc. Natl. Acad. Sci. U.S.A.

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Microsomal preparations from the stalk median eminence of female rats are shown to contain an enzymic activity that is responsible for the formation of MSH-release-inhibiting factor (MSH-R-IF). The amount of this activity remains constant throughout the estrous cycle. The corresponding mitochondrial preparations from the stalk median eminence contain another enzymic principle, estrous cycle-dependent, which competes with the enzyme present in the microsomal preparation for the same "substrate", and can thereby prevent the formation of MSH-R-IF.Several neurohypophyseal hormones, analogs, and peptide intermediates have been tested for their intrinsic MSH-R-IF activity and for their ability to be transformed into MSH-R-IF by incubation with microsomal preparations of stalk median eminence from male rats; it is concluded that the enzyme responsible for the formation of MSH-R-IF is an exopeptidase and that the release-inhibiting factor itself is a tripeptide. Oxytocin is converted by the incubation to L-prolyl-L-leucylglycinamide; nanogram amounts of this tripeptide inhibit the release of MSH from the pituitary both in vivo and in vitro.Rat hypothalamus contains a factor (MSH-R-IF) that inhibits the release of melanocyte-stimulating hormone (1, 2). Evidence for the enzymic formation of this factor has been presented (3). In male rats the pituitary content of MSH remains constant, whereas in females it fluctuates as a function of the estrous cycle (4).This last-mentioned finding was the point of departure for investigating the manner in which pituitary MSH content reflects a change in the enzymic activities responsible for the control of MSH-R-IF formation. We show that a microsomal exopeptidase in the hypothalamus degrades oxytocin, liberating the hormonal C-terminal tripeptide, prolyl-leucylThis work has been performed in C6rdoba and New York. Therefore, different strains of rats and of toads (Bufo arenarum, Bufo marinus) were employed. Since the results obtained in the two laboratories showed no discrepancies, combined publication was agreed upon.Abbreviations of amino-acid derivatives and peptides are in accordance with the IUPAC-IUB Tentative Rules on Biochemical Nomenclature, Biochemistry, 5, 2485Biochemistry, 5, , 1445Biochemistry, 5, (1966 6, 362 (1967

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Hormone Therapeutics – Systemic Hormonal Preparations, Excluding Reproductive Hormones and Insulins (H)

Weinberger

¹

2017

Ullmann's Encyclopedia of Industrial Chemistry

0

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The article contains sections titled: 1. Introduction 2. H01 Pituitary and Hypothalamic Hormones and Analogs 2.1. H01A Anterior Pituitary Lobe Hormones and Analogs 2.1.1. H01AA Adrenocorticotropic Hormone 2.1.2. H01AB Thyrotropin 2.1.3. H01AC Somatropin and Somatropin Agonists 2.1.4. H01AX Other Anterior Pituitary Lobe Hormones and Analogs 2.2. H01B Posterior Pituitary Lobe Hormones 2.2.1. H01BA Vasopressin and Analogs 2.2.2. H01BB Oxytocin and Analogs 2.3. H01C Hypothalamic Hormones 2.3.1. H01CA Gonadotropin‐Releasing Hormones 2.3.2. H01CB Somatostatin and Analogs 2.3.3. H01CC Anti‐Gonadotropin‐Releasing Hormones 3. H02 Corticosteroids Systemic 3.1. H02A Corticosteroids for Systemic Use, Plain 3.1.1. H02AA Mineralocorticoids 3.1.2. H02AB Glucocorticoids 4. H03 Thyroid Therapy 4.1. H03A Thyroid Preparations 4.1.1. H03AA Thyroid Hormones 4.2. H03B Antithyroid Preparations 4.2.1. H03BA Thiouracils 4.2.2. H03BB Sulfur‐Containing Imidazole Derivatives 4.2.3. H03BX Other Antithyroid Preparations 5. H04 Pancreatic Hormones 5.1. H04AA Glycogenolytic Hormones 6. H05 Calcium Homeostasis 6.1. H05AA Parathyroid Hormones and Analogs 6.2. H05B Anti‐Parathyroid Agents 6.2.1. H05BA Calcitonin Preparations 6.2.2. H05BX Other Anti‐Parathyroid Agents

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A solid-phase synthesis of [lysine] vasopressin through a crystalline protected nonapeptide intermediate

Cited by 38 publications

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Solid-Phase Synthesis with Attachment of Peptide to Resin through an Amino Acid Side Chain: [8-Lysine]-Vasopressin

Solid-Phase Synthesis with Attachment of Peptide to Resin through an Amino Acid Side Chain: [8-Lysine]-Vasopressin

Regulation of Formation and Proposed Structure of the Factor Inhibiting the Release of Melanocyte-Stimulating Hormone

Hormone Therapeutics – Systemic Hormonal Preparations, Excluding Reproductive Hormones and Insulins (H)

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