Bcl2 Inhibitor and DNA Hypomethylating Agent Prolong Survival in Patient-Derived Xenograft Model of Down Syndrome Myeloid Leukemia By Synergistic Downregulation of Cytokine Signaling

Background: Down syndrome is known for its leukemia predisposition effects. Children with Down syndrome myeloid leukemia who fail chemotherapy, have a poor outcome. No targeted therapies are available for this population. We have generated and extensively characterized several patient-derived xenograft (PDX) models of Down syndrome AML (Barwe et al., 134:2683, Blood, 2019). We evaluated the efficacy of a combination of DNA hypomethylating agent, azacitidine and Bcl2 inhibitor, venetoclax in PDX model of Down syndrome AML. Although this drug combination is used in the clinic for the treatment of adult AML, their combinatorial mechanism of action is not well known. Methods: NTPL-386 (3 x 10 6 cells) were injected intravenously in NSG-SGM3 mice. Disease progression was monitored by determination of the percentage of human chimerism in peripheral blood by flow cytometry. Mice were randomly assigned to treatment groups (n=5) on day 13 when human CD45+ cells were detectable in blood. Azacitidine (2.5 mg/Kg) was administered intraperitoneally on days 13-19 and venetoclax (100 mg/Kg) was delivered orally on days 13-40 (Fig. 1A). Mice were monitored daily for pre-determined experimental endpoints and were euthanized when any of the endpoints were attained. Kaplan-Meier survival plots were generated. AML cells were harvested from treated mice and lysed, RNA was isolated and sequenced. Pathway analysis was performed to identify the mechanism of action of these two drugs in combination. Results: Azacitidine treatment for one week increased median survival by 8 days compared to untreated mice (Fig. 1B). Venetoclax treatment for 4 weeks was more effective than azacitidine and extended survival by 27 days (P<0.005, compared to untreated). We observed that the combination of venetoclax and azacitidine was much more efficient and prolonged survival by 39 days in this PDX model of refractory Down syndrome AML (P<0.005 and P<0.05 compared to single agent azacitidine or venetoclax respectively). This combination performed better than chemotherapy consisting of cytarabine and daunorubicin, which showed a 11-day improvement in median survival compared to untreated in our previous study (Barwe et al., 134:2683, Blood, 2019). To understand the mechanism of synergism between these two drugs, we conducted transcriptome analysis on cells harvested from mice (n=3 per group) receiving venetoclax and azacitidine, alone or in combination. The log fold changes for each treatment with respect to untreated were calculated and filtered based on FDR>0.05. The genes that were differentially regulated in the combination treatment (Fig. 1C, blue circle in the Venn diagram) were subjected to ToppGene analysis. A heatmap of the differentially regulated genes shows clustering of the replicates from individual mice (Fig. 1D). 'Hematopoietic cell lineage' and 'cytokine signaling in immune system' were the top modulated pathways in the combination treatment. A heatmap of the logFC of the differentially regulated genes from this pathway showed the modulation of these genes in the combination in comparison with individual drug treatment (Fig. 1E). The differentially regulated genes in the combination treatment that showed high variance from the range defined by individual drug treatment belonged to one of three categories - cell surface proteins, cytokines, and stem cell markers. These data indicate that the synergistic downregulation of cytokine signaling is likely responsible for the combinatorial effect of venetoclax and azacitidine. Conclusion: The combination of venetoclax and azacitidine was effective in prolonging survival in a highly refractory Down syndrome AML PDX model (NTPL-386) generated in the laboratory. The top genes differentially regulated by the combination treatment which varied the most in magnitude from the individual drug treatment identified genes that are likely to contribute to the synergism between the two drugs. Figure 1 Figure 1. Disclosures Barwe: Prelude Therapeutics: Research Funding. Gopalakrishnapillai: Geron: Research Funding.

Short term treatment with a cocktail of rapamycin, acarbose and phenylbutyrate slows aging in mice

Pharmaceutical intervention of aging requires targeting multiple pathways, thus there is rationale to test combinations of drugs each targeting different but overlapping processes. In order to determine if combining drugs previously shown to improve lifespan would have greater impact than any individual drug, a diet containing rapamycin at 14 ppm, acarbose at 1000 ppm, and phenylbutyrate at 1000 ppm was fed to 20-month-old C57BL/6 and HET3 4-way cross mice of both sexes for three months. Mice fed the cocktail diet showed a strain and gender-dependent phenotype consistent with healthy aging including decreased body fat and blood glucose, improved cognition, and increased grip strength and walking ability compared to mice fed individual drug or control diets. A cocktail diet containing one-half dosing of each compound was overall less effective than the full dose. The composite age-related lesion score of heart, lungs, liver and kidney was decreased in mice fed the cocktail diet compared to mice fed individual drug or control diets suggesting an interactive advantage of the three drugs. Senescence and inflammatory cytokine levels in kidneys from mice fed the cocktail diet were lower than in kidneys from mice fed control diet, and consistent with low expression levels in kidneys from young untreated mice, suggesting the cocktail diet delayed aging partly by senolytic and anti-inflammatory effects.

Analysis of the variations in price of anti-glaucoma eye preparations available in Indian pharmaceutical market

Background: The aim of the study was to analyze the percentage cost variations among different brands of the commonly prescribed anti–glaucoma drugs.Methods: The maximum and minimum price of each brand of the drug in INR was noted by using CIMS January to April 2018 edition Drug Today April to June 2018 Vol-1. The cost ratio and the percentage cost variation for individual drug brands was calculated. The cost of each eye drop was calculated. At last the cost ratio and percentage cost variation of various brands was compared.Results: Percentage variation in cost for anti-glaucoma eye preparations marketed in india was found to be eye drop timolol maleate (0.5%) of 5 ml:263.63, eye drop dorzolamide (2%) of 5 ml:9.77, eye drop pilocarpine (2%) of 5 ml:160.40, eye drop Betaxolol (0.5%) of 5 ml:56.54, eye drop Latanoprost (50 mcg/ml) of 2.5ml:135.88, eye drop Brimonidine tartarate (0.15%) of 5 ml:183.9, eye drop Levobunolol (5 mg/ml) of 5 ml:32.38.Conclusions: Glaucoma is the most common ocular disease and eye drops are to be prescribed for prolonged period. If a costly brand is prescribed, the patients have to pay more money unnecessarily for their treatment. The clinicians prescribing these drugs should be aware of these variations in cost to reduce the cost of drug therapy.

Potential Therapeutic effect of Bee Venom on Cisplatin-Induced Hepatotoxicity

The fact that cisplatin (CIS) can induce hepatotoxicity has limited its therapeutic uses, although it is recognised as a highly potent antineoplastic drug. Bee venom (BV) is an important toxin that exerts a potent anti-inflammatory effect and demonstrates significant pharmacological activities. The aim of this research was to investigate the therapeutic impact of BV administration orally on CIS-induced hepatotoxicity in a rat model when compared with individual drug therapy. Thirty albino male rats were used in this study. The rats were classified into three groups, each with ten rats (n = 10). The control group was group I, while groups II and III got CIS (a single dose of 7.5 mg/kg interperitoneally). After 24 hours, group III received BV (1 mg/kg orally), which was then administered daily for four weeks. The rats’ serum liver functions were estimated. The oxidative stress, antioxidant status, inflammatory mediators and fibrogenic and apoptotic markers were determined in the liver tissues. The results revealed that the administration of CIS induced a significant elevation in the serum aspartate aminotransferase, alanine aminotransferase and bilirubin levels, which was associated with a decrease in the albumin level. Additionally, significant increases in the hepatic tissue malondialdehyde, tumor necrosis factor alpha, interferon gamma, transforming growth factor beta, fibronectin and caspase-3 levels were noted. Moreover, significant decreases in the superoxide dismutase and catalase activities as well as the glutathione level were detected in the hepatic tissues. The administration of BV resulted in the notable amelioration of the aforementioned parameters. Interestingly, these parameters were restored to almost normal levels following administration of CIS and BV. The histopathological investigation revealed multiple focal inflammation sites that appeared to be associated with hepatic necrosis in the hepatic tissues of the CIS-treated rats. The co-administration of BV preserved the normal architecture of the hepatic tissues. These results indicate that the co-administration of BV exerts potent anti-hepatotoxic and cytoprotective effects by combating oxidative stress, inflammation, fibrogenic and apoptotic markers and histopathological changes. Thus, this study provides strong evidence of the superiority of combination drug therapy when compared with individual drug therapy.

Drug Use Disorders and Violence: Associations With Individual Drug Categories

We conducted a systematic review that examined the link between individual drug categories and violent outcomes. We searched for primary case-control and cohort investigations that reported risk of violence against others among individuals diagnosed with drug use disorders using validated clinical criteria, following Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. We identified 18 studies published during 1990–2019, reporting data from 591,411 individuals with drug use disorders. We reported odds ratios of the violence risk in different categories of drug use disorders compared with those without. We found odds ratios ranging from 0.8 to 25.0 for most individual drug categories, with generally higher odds ratios among individuals with polydrug use disorders. In addition, we explored sources of between-study heterogeneity by subgroup and meta-regression analyses. Cohort investigations reported a lower risk of violence than case-control reports (odds ratio = 2.7 (95% confidence interval (CI): 2.1, 3.5) vs. 6.6 (95% CI: 5.1, 8.6)), and associations were stronger when the outcome was any violence rather than intimate partner violence (odds ratio = 5.7 (95% CI: 3.8, 8.6) vs. 1.7 (95% CI: 1.4, 2.1)), which was consistent with results from the meta-regression. Overall, these findings highlight the potential impact of preventing and treating drug use disorders on reducing violence risk and associated morbidities.