Determination of the configuration of histrelin,and LHRH analog,by chiral capillary gas chromatography

Summary Summary The assigned chirality at each center of the synthetic nonapeptide histrelin (L-pyroglutamyl-L-histidyl-L-tryptophyl-L-seryl-L-tyrosyl-D-N^im-benzyl-histidyl-L-leucyl-L-arginyl-L-proline-ethylamide) was verified using chiral gas chromatography. The procedure involved acid hydrolysis of histrelin to the constituent amino acids, derivatization as the N-pentafluoropropionyl/isopropyl esters and the analysis of the mixture using a commercially available 25m chiral capillary column (Chirasil-L-Val). There was no significant difference in the retention time of the amino acids obtained from the hydrolysate mixture when compared to the appropriate standards. Additionally, the hydrolysate was spiked with the D and L amino acids to prove the identity of closely eluting peaks. Luteinizing hormone-releasing hormone     

A new strategy utilizing electrospray ionization-quadrupole ion trap mass spectrometry for the qualitative determination of GnRH peptides

Numerous forms of the neurotransmitter GnRH have been discovered in vertebrates and invertebrates. Methods used for identification of these peptides are laborious and often require the application of multiple, confirmatory techniques. In this study, we investigate whether HPLC-MS/MS and de novo sequencing techniques applied to whole peptide analysis can provide a simpler approach to GnRH characterization. Experiments were performed with six GnRH forms (chicken I, chicken II, lamprey III, mammalian, salmon and seabream) to determine whether MS/MS spectra would be dominated by proline-directed fragmentation to the detriment of obtaining sufficient fragmentation for sequencing. While the expected b8 fragment was prominent, sufficient ion series were obtained for the six GnRH peptides to provide sequence identification. On the basis of the patterns observed for six model peptides, similar fragmentation patterns are expected for other GnRH forms. To confirm the applicability of the method, extracts from Sprague-Dawley rat brains were examined. These experiments confirm the presence of mammalian GnRH and a posttranslationally modified form of mammalian GnRH, hydroxyproline9 GnRH, in Sprague-Dawley rat brains and demonstrate that ESI-MS/MS techniques provide a valuable addition to existing qualitative methods.     

长效LHRH拮抗剂的设计、合成和生物活性

根据抗蛋白酶降解的长效肽设计思想, 合成了一系列新型结构的LHRH拮抗剂类似物. 体内生物活性评价结果表明, 所设计的多肽具有比母体肽和阳性对照更长的体内抑制睾酮作用时间和较低的最低有效剂量, 证实了该设计思想的可行性, 并为开发长效LHRH拮抗剂药物提供了新的候选化合物.     

Synthesis of a β-turn mimetic suitable for incorporation in the peptide hormone LHRH

LHRH is a decapeptide hormone which plays a central role in neuroendocrinology. Conformational studies have suggested that LHRH may adopt a β-turn involving residues 5-8 when bound to its receptor. A β-turn mimetic with side chains corresponding to those of a Tyr-Gly-Leu-Orn tetrapeptide has therefore been synthesized for incorporation at positions 5-8 in LHRH. In the turn mimetic, residues i and i+1 are connected by a ψ[CH₂O] isostere instead of an amide bond, while a covalent ethylene bridge replaces the hydrogen bond which is often found between residues i and i+3 in β-turns. The turn mimetic was assembled from three types of building blocks: an azido aldehyde, an Fmoc protected amino acid and a protected dipeptide amine.     

EFFECTS OF CHANGES IN PATTERN OF LHRH PULSE ON LH SECRETION BY PERFUSED ANTERIOR PITUITARY CELLS OF MALE RATS

In the present study, a perfusion system of dispersed cells was used to investigate theeffects of LHRH pulse amplitude and frequency, and LHRH continuous stimulation on LHsecretion by anterior pituitary cells of adult male rats. The results have shown that, in therange of LHRH concentrations from 1×10~(-10) to 1 ×10~(-6) mol/L, the dose-response curve of LHsecretion was linear. LHRH pulse frequency generated a biphasic LH response: increasingLHRH pulse frequency increased the basal LH secretion and decreased LH/pulse. When1 ×10~(-9), mol/L or greater LHRH was given at frequencies of 3 pulses/h or higher, it wasobserved that a maximal LH peak was induced and then the LH release declined progres-sively to its LH basal level, i.e. LHRH self-priming effect and LH desensitization occurred.Enhancement of amplitude of LHRH pulses could reduce pulse frequency required for prim-ing. Increases in frequency of LHRH pulses with high amplitude would provoke the prim-ing effect more quickly. In addition, continuou     

Chemistry and Molecular Biology in the Search for New LHRH Antagonists

Hormones—and in particular, the sex hormones—were the first growth factors discovered to be involuntary helpers of cancer. Female breast cancer and male prostate cancer are the best known examples of tumors acknowledged to be hormone-dependent. A look at cancer statistics shows that breast cancer is still the most frequent cancer in women; in men, prostate cancer plays a similarly dominant role with increasing age. Shutting down the main production site of the sex hormones estrogen and testosterone either by removing the ovaries or by castration is a well-known and often effective therapy; however, these procedures can be problematic due to the concomittant psychological stress. Modern hormone therapy for advanced breast cancer and prostate cancer attempts to spare the patient such irreversible operative procedures for as long as possible, by using hormone antagonists, such as the LHRH antagonists, which hinder deployment of the hormone itself and thus its growth-promoting activity.     

Expression of Luteinizing Hormone-Releasing Hormone (LHRH) and Type-I LHRH Receptor in Transitional Cell Carcinoma Type of Human Bladder Cancer

Bladder cancer (BC) is the tenth most frequently detected cancer in both sexes. Type-I luteinizing hormone-releasing hormone (LHRH) receptor (LHRH-R-I) is expressed not only in the pituitary, but also in several types of cancer disease. There are few data about LHRH-R-I expression in human BC. This study aimed to investigate the expression of LHRH and LHRH-R-I in the transitional cell carcinoma (TCC) type of human BC. RNA was extracted from 24 human bladder tumor specimens and three BC cell lines. RT-PCR was performed to detect mRNA for LHRH and LHRH-R-I. The protein of LHRH-R-I was further studied by immunohistochemistry (IHC), ligand competition assay, and Western Blot. PCR products of LHRH were found in 19 of 24 (79%) specimens and mRNA of LHRH-R-I was detected in 20 of 24 specimens (83%). Positive immunostaining for LHRH-R-I with different expression intensity was found in all samples examined, showing negative correlation with TCC grade. Radioligand binding studies also showed the presence of specific LHRH-R-I and high affinity binding of LHRH analogs. The high incidence of LHRH-R in BC suggests that it could serve as a molecular target for therapy of human BC with cytotoxic LHRH analogs or modern powerful antagonistic analogs of LHRH.     

Comment on “d-Poly(lactide) and LHRH decapeptide stereointeractions investigated by vibrational spectroscopy”

This note is a response to Zhang et al. [Zhang J, Beshra A, Domb AJ, Ozaki Y. d-Poly(lactide) and LHRH decapeptide stereointeractions investigated by vibrational spectroscopy. Eur Polym J 2007;43:3016-27] who reported spectroscopic data for blends of poly-d-lactide and LHRH (a leuprolide decapeptide analog). These authors concluded to the presence of a stereocomplex between those two species but it is shown, in this note, that the results obtained as well as literature results do not support this conclusion.     

甲酸铵催化转移氢化还原肽链中的芳香硝基——对氨基苯丙氨酸的间接引入

研究了用甲酸铵催化转移氢化法(AF-CTH)对不同类型肽中的芳香硝基的还原行为, 这些肽类化合物包括促黑激素(MSH: 四肽)、促黄体素释放激素(LHRH: 十肽)和强腓肽(十七肽)的类似物. 用HPLC对还原过程进行了跟踪监测, 结果显示, 除含对氯苯丙氨酸残基的LHRH类似物因发生脱氯副反应不适合用AF-CTH还原外, 其余序列还原过程中均无明显副反应发生, 硝基几乎定量地转化成为相应的氨基, 实现了对氨基苯丙氨酸向肽链的间接引入. 另外发现, 硝基还原所需的时间与肽链长度有关, 肽链越长, 还原所需时间越长, 但与其在序列中的位置关系不明显.     

SYNTHESIS AND BIOACTIVITIES OF NEW LHRH ANTAGONISTS CONTAINING NOVEL UNNATURAL AMINO ACIDS AT POSITION FIVE

Synthesis and bioactivities of new antagonists of luteinizing hormone releasing hormone(LHRH) with novel unnatural amino acids at position five are reported, Most of them showedsome antiovulatory activity at 0. 5μg/rat and two of them inhibited ovulation completely at1μg/rat using saline as vehicle.