Mohamed Alaama

Anticancer Effects of Medical Malaysian Leech Saliva Extract (LSE)

Leech saliva contains biologically active compounds that are mainly proteins & peptides. Small cell lung cancer (SCLC) is a form of cell lung carcinoma. In this study a modified and smooth extraction method of saliva was used without leech scarification. Trying to find out the biological activity of medical Malaysian Leech Saliva Extract as cytotoxic in vitro, the SW 1271 cell line was grown and maintained in Leibovitzs L-15 medium supplemented with 10% foetal bovine serum at 37 ° C in normal atmospheric air. Serial dilutions of LSE were added to the cell line SW 1271 media for testing the cytotoxic activities.Result revealed that the LSE has a cytotoxic activity against small cell lung cancer(SW 1271 cell line) with IC 50 of 119.844 µg/ml compared with IC 50 values of two reference standard drugs Irinotecan (5.81 µg/ml) and Carboplatin(18.754 µg/ml). In a combination regimen, LSE reduced the IC 50 of Carboplatin & Irinotecan by 65% & 11.5% respectively. Carboplatin reduced the IC 50 of LSE by 4.6%, while Irinotecan reduced it by 57%. This results provides a promising novel agent for treatment of small cell lung cancer (SCLC) at least in vitro, more researches are needed.

Abstract 5130: Assessment of the antitumor activity of leech (huridinaria manillensis) saliva extract in prostate cancer

BACKGROUND Ancient traditional physicians from many countries used leeching to treat a wide range of diseases for thousands of years. Leech saliva contains a large number of peptides and proteins, which have anti-thrombotic, antimicrobial, antitumor and anti-metastatic activities. Currently, leech therapy has an established role as an important tool in microsurgery, reconstructive surgery and salvage of grafted tissues. METHODS Leech saliva extract (LSE) was prepared as an aqueous solution from frozen lyophilized powder. LSE IC50 was determined in-vitro in five prostate cancer cell lines using MTS cell viability assay. In-vivo efficacy of LSE was determined in LNCaP and 22RV-1 in nude mice xenograft models. Mice were injected with 2×106 LNCaP or 22RV-1 cells subcutaneously; the mice were castrated in both studies to resemble castration resistant prostate cancer. After castration, mice were divided into four groups of 6-8 each. Mice were subcutaneously injected with either LSE (5 mg/kg) once a week, LSE (5 mg/kg) twice a week, docetaxel (10 mg/kg) or vehicle twice a week. PSA, tumor volume, and weight were measured weekly in the LNCaP model; and in the 22RV-1 tumor volume and weights were measured twice weekly. After four weeks of treatment, mice were euthanized, tumors and organs were collected for transcriptome and histopathological analysis. RESULTS LSE induced cell death in a panel of prostate cancer cell lines including LNCaP, PC3 and 22RV-1. IC50 values of were determined as 22 μg/ml in both LNCaP and PC3 cells and 53 μg/ml in 22RV-1cells. Furthermore, in vivo studies show that LSE once and twice weekly regimens both caused significant decrease in PSA and tumor volume compared to control. There was no significant difference between the antitumor activity of LSE and docetaxel. Interestingly, once weekly treatment with LSE was associated with significant weight gain (due to good dietary intake) at several time points. Immunohistochemical staining (IHC) showed significant increase in caspase-3 and significant decrease in P21, Ki-67, and PCNA expression in the LSE treated mice compared to the control group. Transcriptome analysis of tumor samples showed that LSE had significant immunomodulatory, anti-inflammatory, along with significant effects on cell-cell adhesion, induction of glutathione transferase and inhibition of certain growth factors. Consequently, these effects led to significant cell cycle arrest, increase in apoptosis and decrease in proliferation. CONCLUSIONS LSE has significant in vitro and in vivo anti-tumor activity with no apparent side effects. This can be attributed, at least partly, to its ability to inhibit cellular proliferation and induce apoptosis through its modulation of immunity, cell-cell adhesion, and inflammation. Citation Format: Amr E. Ammar, Mohamed H. Hassona, Gray R. Meckling, Leslie G. Chan, Mei Y. Chin, Abdulrahman Abdualkader, Mohamed Alaama, Ahmed Merzouk, Abulbashar Helaluddin, Abbas Ghawi, Omer Kucuk, Emma S. Guns. Assessment of the antitumor activity of leech (huridinaria manillensis) saliva extract in prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5130. doi:10.1158/1538-7445.AM2015-5130

Free radical scavenging activity of the Malaysian Leech Saliva extract, Hirudinaria manillensis

D discovery is a complex process which involves an interdisciplinary approach to design effective feasible drugs (1). The development of new drugs with potential therapeutic applications is one of the most complex process in the pharmaceutical industry (2). Millions of dollars and man hours are dedicated to the discovery of new therapeutic agents. Rational drug discovery process combat and supersede the conventional process with the advent of proteomic, genomic and structural information (3).U in conjunction with other technologies the light beam becomes a uniquely powerful tool to study cells. Currently light source is used not only to observe cells, but also to stimulate cellular chemistry. This area of biophotonics is called “caged compounds” as synthetic organic chemistry say “Photolabile compounds” is used to make biologically signalling molecules functionally inert. Irradiation with light liberates the caged molecules, thus “switching on” a chosen signaling pathway. Such light-triggered release of molecules from “caged” forms also offers the potential to deliver innocuous agents to cells, tissues, and organisms, where they can be unmasked to their active states. Because light can be readily tuned and focused, it can be spatially and temporally controlled to provide “on-command” drug delivery, unmasking of biochemical agents for enzyme and protein activation, and other biochemical and physiological studies.T effects of artemisinin derivatives, artemether and lumefantrine on the activity of glucose 6-phosphate dehydrogenase (G6PD) and some haematological parameters in rats (Rattus novergicus) were studied. The experimental animals were randomly distributed into four groups: those administered Tween 80 (control), those administered artemether (8 mg/kg body weight), those administered lumefantrine (48 mg/kg body weight) and those co-administered artemether (8 mg/kg body weight) and lumefantrine (48 mg/kg body weight). The drugs were orally administered twice daily for three days to half of the animals while the remaining half were dosed for six days. Animals were subsequently anaesthetized in diethyl ether, blood samples were collected by cardiac puncture and the organs were excised and weighed. The following parameters were assessed in blood and liver homogenate: glucose, G6PDH, packed cell volume (PCV), haemoglobin (HB), white blood cell count (WBC), neutrophil, leukocyte, eosinophil, monocyte and basophil. After 3 days of administration, results showed significant decreases (p 0.05) in plasma and homogenate glucose, G6PD activity and haematological parameters. It could be concluded that the administration of artemether and lumefantrine after 3 days to non-malaria infected rats showed hypoglycaemia and reduced haematological indices, while increasing G6PD activity.T study is to reveal whether artificial hemolyzed blood samples obtained during clinical operation affect the pharmacokinetic (PK) profile and bioequivalence (BE) study results. A validated LC-MS/MS method was applied to analyze both hemolyzed and non-hemolyzed plasma samples derived from healthy volunteers to whom clopidogrel, methylprednisolone and ropinirole were administrated orally in three different pilot BE studies conducted in the clinical research center. During the first period of BE studies, for drug concentrations of hemolyzed and non-hemolyzed plasma samples, clopidogrel were 862.57±860.16 (pg/mL) and 920.61±959.14 (pg/mL) at 1 hour post dosing; methylprednisolone were 155.21±33.60 (pg/mL) and 160.01±29.9 (pg/mL) at 2.5 hours post dosing; ropinirole were 1322.87±392.96 (pg/mL) and 1151.42±299.91 (pg/mL) at 4 hours post dosing. During the second period, the values are 895.61±590.47 (pg/mL) and 941.60±601.91 (pg/mL) for clopidogrel; 160.01±29.99 (pg/mL) and 127.40±41.61 (pg/mL) for methylprednisolone; 1146.30±249.89 (pg/mL) and 1220.01±196.67 (pg/mL) for ropinirole. The drug concentration between hemolyzed and non-hemolyzed plasma samples did not yield a significant difference (p>0.05). In conclusion, although hemolysis may physically change the characteristics of the plasma samples, it doesn’t significantly affet the accuracy of PK profile of clopidogrel, methylprednisolone and ropinirole. However, hemolysis should always be avoided in the practice of clinical studies to get ideal plasma for analysis.Free radical scavenging activity of the Malaysian Leech Saliva extract, Hirudinaria manillensis Abbas Mohammad Ghawi1, Abdualrahman M.Abdualkader2, Ahmed Merzouk3 and Mohamed Alaama2 Basic Medical Science Department, Faculty of Pharmacy, International Islamic University Malaysia, Malaysia. Pharmaceutical Chemistry Department, Faculty of Pharmacy, International Islamic University Malaysia, Malaysia BIOPEP SOLUTIONS INC., Vancouver, BC CanadaTrypanosomatids cause deadly diseases in humans. Of the various biochemical pathways in trypanosomatids, glycolysis, has received special attention because of being sequestered in peroxisome like organelles critical for the survival of the parasites. This study focuses on phosphoglycerate kinase (PGK) from Leishmania spp. which, exists in two isoforms, the cytoplasmic PGKB and glycosomal PGKC differing in their biochemical properties. Computational analysis predicted the likelihood of a transmembrane helix only in the glycosomal isoform PGKC, of approximate length 20 residues in the 62-residue extension, ending at, arginine residues R471 and R472. From experimental studies using circular dichroism and NMR with deuterated sodium dodecyl sulfate, we find that the transmembrane helix spans residues 448 +/- 2 to 476 in Leishmania mexicana PGKC. The significance of this observation is discussed in the context of glycosomal transport and substrate tunneling

A Review on the Formulation and Analysis of Anti-Diabetic Agent: Gliclazide

Gliclazide is a second generation sulfonylurea, which is used as antidiabetics drug. It is orally administrated and used for the treatment of non-insulin-dependent diabetes mellitus (NIDDM) and has duration of action of 12 h or more. Beside its hypoglycaemic effects, gliclazide was reported to have many other important effects, such as: suppression of platelet functions, antithrombotic actions, decreased the production of tumour necrosis factor (TNF) α by endothelial cells and other effects. The unique activities of gliclazide has open a new avenue for the drug development research. This work is aimed to provide comprehensive information about gliclazide and its current research activities.