Antiretroviral Agents Research Articles

Plasma Levels of Secretory Calreticulin among HIV Infected Patients in Bauchi State

Objectives: Human immunodeficiency virus (HIV) is a lentivirus that causes HIV infection and over time acquired immunodeficiency syndrome. The objective was to evaluate plasma levels of Calreticulin (CALR) in HIV positive and negative individuals. Methods: A total of 164 participants were recruited and divided into four groups according to their viral load after estimation and individuals recruited as controls. Viral load was estimated in 1.1 ml of plasma according to manufacturers (COBAS Taqman HIV-1 Quantitative Test, v2.0, Roche Diagnostic GmbH, Germany). Plasma levels of CALR were analysed using ELISA (ELISA-Biotuva Life Sciences, UK). Results: Result revealed a significant difference (p = 0.00) between the CALR levels of individuals with low viral load and individuals with unsuppressed viral load, suppressed viral load, and controls. A significantly (p = 0.017) higher plasma CALR in individuals with unsuppressed viral load compared to those with suppressed viral load was found. No differences between plasma CALR levels of individuals with unsuppressed viral load and that of controls. Conclusion: Our study showed up regulation of plasma CALR in HIV-infected patients on long term antiretroviral therapy. We believe and from existing literature this up regulation is due partly if not solely to the oxidative stress and endoplasmic reticulum stress caused by the antiretroviral agents.

Structure-Based Design of Tropane Derivatives as a Novel Series of CCR5 Antagonists with Broad-Spectrum Anti-HIV-1 Activities and Improved Oral Bioavailability.

Blocking the entry of an HIV-1 targeting CCR5 coreceptor has emerged as an attractive strategy to develop HIV therapeutics. Maraviroc is the only CCR5 antagonist approved by FDA; however, serious side effects limited its clinical use. Herein, 21 novel tropane derivatives (6-26) were designed and synthesized based on the CCR5-maraviroc complex structure. Among them, compounds 25 and 26 had comparable activity to maraviroc and presented more potent inhibitory activity against a series of HIV-1 strains. In addition, compound 26 exhibited synergistic or additive antiviral effects in combination with other antiretroviral agents. Compared to maraviroc, both 25 and 26 displayed higher Cmax and AUC0-∞ and improved oral bioavailability in SD rats. In addition, compounds 25 and 26 showed no significant CYP450 inhibition and showed a novel binding mode with CCR5 different from that of maraviroc-CCR5. In summary, compounds 25 and 26 are promising drug candidates for the treatment of HIV-1 infection.

In Vitro Susceptibility of HIV Isolates with High Growth Capability to Antiretroviral Drugs

It has been considered that reduced susceptibility to antiretroviral drugs is influenced by drug adherence, drug tolerance and drug-resistance-related mutations in the HIV genome. In the present study, we assessed the intrinsic high viral growth capability as a potential viral factor that may influence their susceptibility to antiretroviral drugs using an in vitro model. Phytohemagglutinin-activated peripheral blood mononuclear cells (1.5 × 106 cells) were infected with HIV isolates (106 copies/mL). The culture was carried out at different concentrations (0.001-20 μM) of 13 synthetic antiretroviral compounds (six nucleoside/nucleotide reverse transcriptase inhibitors, one non-nucleoside reverse transcriptase inhibitor, four integrase inhibitors, and two protease inhibitors), and HIV production was assessed using HIV-RNA copies in culture. The 90% inhibitory concentration (IC90) and pharmacokinetics of an antiretroviral agent were used as parameters to determine the reduced antiretroviral drug susceptibility of HIV isolates with high growth capability to synthetic antiretroviral compounds. The high growth capability of HIV isolates without any known drug resistance-related mutation affected their susceptibility to tenofovir (IC90 = 2.05 ± 0.40 μM), lamivudine (IC90 = 6.83 ± 3.96 μM), emtricitabine (IC90 = 0.68 ± 0.37 μM), and efavirenz (IC90 = 3.65 ± 0.77 μM). These antiretroviral drugs showed IC90 values close to or above the maximum plasma concentration against HIV isolates with high growth capability without any known drug resistance-related mutation. Our results may contribute to the development of effective strategies to tailor and individualize antiretroviral therapy in patients harboring HIV isolates with high growth capability.

An evaluation of postmarketing reports of hyperglycaemia associated with dolutegravir for treatment of HIV in Eswatini

BackgroundDolutegravir (DTG) is an Integrase Strand Transfer Inhibitor (INSTI) indicated in combination with other antiretroviral agents for the treatment of HIV infection. It is available in a number of pharmaceutical preparations including the fixed-dose combination (TLD) containing tenofovir (300 mg) + lamivudine (300 mg) + dolutegravir (50 mg). In 2018, Eswatini adopted TLD as the preferred first-line HIV treatment regimen for adults and adolescents as per WHO recommendations. From March 2019 to March 2020, the National Pharmacovigilance Center (NPC) in Eswatini received 8 reports of hyperglycaemia associated with the use of DTG. This study was conducted to investigate if Eswatini NPC database included cases suggestive of causality between dolutegravir and hyperglycaemia.MethodA qualitative synthesis of information from the Eswatini national pharmacovigilance database from March 2019 to March 2020 was conducted to investigate a casual association between hyperglycaemia and dolutegravir.ResultsAll reports with dolutegravir containing regimen and suspected Adverse Event of hyperglycaemia in the period of March 2019 to March 2020 were included in the study. Seven of the reports were serious (resulted in hospitalization and one case concerned optic neuritis, leading to blindness). Two patients had a medical history of diabetes while the rest of the patients had never experienced hyperglycaemia before starting dolutegravir. For all the reports, the time to onset of hyperglycaemia ranges from 2–5 months after the initiation of DTG. None of the patients discontinued the use of DTG. All the patients were treated with oral hypoglycaemic medication. In severe cases, patients were treated with intravenous normal saline and ringer lactate as well as rapid-acting insulins. All patients are currently stable on oral hypoglycaemic drugs.ConclusionCases that support causality between dolutegravir containing regimen and hyperglycaemia were found. These cases were mainly serious. Based on these findings it is recommended that healthcare professionals (HCPs) actively screen all patients for risk factors of hyperglycaemia before DTG initiation. In addition, it is important that HCPs are aware of the possible association between DTG and hyperglycaemia.

Peripheral sensory neuron CB2 cannabinoid receptors are necessary for both CB2-mediated antinociceptive efficacy and sparing of morphine tolerance in a mouse model of anti-retroviral toxic neuropathy

Painful peripheral neuropathy is a common neurological complication associated with human immunodeficiency virus (HIV) infection and anti-retroviral therapy. We characterized the impact of two CB2 cannabinoid agonists (AM1710 and LY2828360 – ligands differing in signaling bias and CNS penetration) on neuropathic nociception induced by the antiretroviral agent Zalcitabine (2',3'-dideoxycytidine; ddC). We also used a conditional knockout approach to identify cell types mediating CB2 agonist-induced antinociceptive efficacy and sparing of morphine tolerance. AM1710 and LY2828360 alleviated ddC-induced neuropathic nociception in mice of both sexes. These benefits were absent in global CB2 knockout mice, which exhibited robust morphine antinociception. Like morphine, AM1710 blunted ddC-induced increases in proinflammatory cytokine (IL-1β, TNF-α) and chemokine (CCL2) mRNA expression levels. We generated advillinCre/+;CB2f/f conditional knockout mice to ascertain the role of CB2 localized to primary sensory neurons in CB2-mediated therapeutic effects. Antinociceptive efficacy of both AM1710 and LY2828360, but not reference analgesics, were absent in advillinCre/+;CB2f/f mice, which exhibited robust ddC-induced neuropathy. In ddC-treated CB2f/f mice, LY2828360 suppressed development of morphine tolerance and reversed established morphine tolerance, albeit with greater efficacy in male compared to female mice. LY2828360 failed to block or reverse morphine tolerance in advillinCre/+;CB2f/f mice. The present studies indicate that CB2 activation may alleviate HIV-associated antiretroviral neuropathy and identify a previously unreported mechanism through which CB2 activation produces antinociceptive efficacy. Our results also provide the first evidence that a CB2 agonist can reverse established morphine tolerance and demonstrate that CB2 localized to peripheral sensory neurons mediates the opioid tolerance sparing efficacy of CB2 agonists.

Dolutegravir potentiates platelet activation by a calcium-dependent, ionophore-like mechanism

Dolutegravir is a highly potent HIV integrase strand transfer inhibitor that is recommended for first-line anti-retroviral treatment in all major treatment guidelines. A recent study has shown that people taking this class of anti-retroviral treatment have a substantially higher risk of early-onset cardiovascular disease, a condition shown previously to be associated with increased platelet reactivity. To date, few studies have explored the effects of dolutegravir on platelet activation. Accordingly, the current study was undertaken with the primary objective of investigating the effects of dolutegravir on the reactivity of human platelets in vitro. Platelet-rich plasma, isolated platelets, or buffy coat cell suspensions prepared from the blood of healthy adults were treated with dolutegravir (2.5–10 µg/ml), followed by activation with adenosine 5’-diphosphate (ADP), thrombin, or a thromboxane A2 receptor agonist U46619. Expression of platelet CD62P (P-selectin), formation of heterotypic neutrophil:platelet aggregates, and calcium (Ca2+) fluxes were measured using flow cytometry and fluorescence spectrometry, respectively. Dolutegravir caused dose-related potentiation of ADP-, thrombin- and U46619-activated expression of CD62P by platelets, as well as a significant increases in formation of neutrophil:platelet aggregates. These effects were paralleled by a spontaneous, receptor-independent elevation in cytosolic Ca2+ that appears to underpin the mechanism by which the antiretroviral agent augments the responsiveness of these cells to ADP, thrombin and U46619. The most likely mechanism of dolutegravir-mediated increases in platelet cytosolic Ca2+ relates to a combination of lipophilicity and divalent/trivalent metal-binding and/or chelating properties of the anti-retroviral agent. These properties are likely to confer ionophore-type activities on dolutegravir that would promote movement of Ca2+ across the plasma membrane, delivering the cation to the cytosol where it would augment Ca2+-dependent intracellular signaling mechanisms. These effects of dolutegravir may lead to hyper-activation of platelets which, if operative in vivo, may contribute to an increased risk for cardiometabolic co-morbidities.

Young age is a key determinant of body weight gain after switching from tenofovir disoproxil fumarate to tenofovir alafenamide in Japanese people living with HIV

BackgroundTreatment with tenofovir alafenamide fumarate (TAF) is associated with body weight gain. However, little or no information is available on this issue in Asian populations. MethodsThis single-center retrospective study included Japanese people living with HIV (PLWH) who satisfied the following criteria; 1) switching from TDF to TAF after HIV-suppression, 2) follow-up for ≥2 years while on TDF and TAF, and 3) no switching of the third antiretroviral agent. Changes in annual body weight and lipid profiles were compared between the TDF and TAF periods. ResultsOf 328 patients, dolutegravir (DTG) was used in 118 PLWH. Overall, no significant difference in weight gain was observed between TDF and TAF (0.76 vs. 0.9 kg/year, p = 0.331). In TAF-period, younger (<50 years of age) group showed significantly greater weight gain than older group (1.03 vs. 0.12 kg/year, p = 0.037). In DTG group, weight gain was larger in TAF-period (0.74 vs. 1.31 kg/year, p = 0.046), especially in younger subgroup (1.43 kg/year) compared with older one (−0.12 kg/year). Multivariate regression analysis showed that TAF was not associated with weight gain (estimates 0.201, p = 0.170) except for DTG group, whereas young age was associated with weight gain in all subjects (estimates −0.033/1 year older, p < 0.001), DTG, RAL, and EFV groups. ConclusionIn Japanese PLWH, annual body weight change was comparable in TDF- and TAF-period, while TAF plus DTG correlated with weight gain. Since young age was a key determinant of weight change, careful interpretation is needed for TAF-associated weight gain.

The risk of hyperglycemia associated with use of dolutegravir among adults living with HIV in Kampala, Uganda: A case-control study.

Emerging evidence suggests a possible association between hyperglycemia and dolutegravir (DTG), a preferred first-line antiretroviral agent in sub-Saharan Africa (SSA). There is need for rigorous studies to validate this association in the face of increasing DTG use and burden of non-communicable diseases among people living with HIV (PLHIV). We conducted a case-control study to assess the risk of hyperglycemia associated with use of DTG among PLHIV attending Mulago ISS Clinic in Kampala. Cases had hyperglycemia while controls had no hyperglycemia as confirmed by fasting plasma glucose and oral glucose tolerance tests. Demographic, laboratory, and clinical data were collected using interviewer-administered questionnaires and medical record abstraction. Analysis compared cases and controls on DTG use prior to diagnosis of hyperglycemia while controlling for potential confounders using multivariable logistic regression. We included 204 cases and 231 controls. In multivariable analysis, patients with prior DTG use had seven times greater odds of subsequent diagnosis of hyperglycemia compared to those who had non-DTG-based regimens (adjusted odds ratio [aOR] 7.01, 95% CI 1.96-25.09). The odds of hyperglycemia also increased with age (56 years and above vs. 18-35, aOR 12.38, 95% CI 3.79-40.50) and hypertension (aOR 5.78, 95% CI 2.53-13.21). Our study demonstrates a strong association between prior DTG exposure and subsequent diagnosis of hyperglycemia. Given the benefits of DTG, wide-scale use, and the growing burden of diabetes mellitus (DM) in SSA, there is need for systematic screening for hyperglycemia and consideration of alternate regimens for those at risk for DM.

IDVIP: identification and characterization of viral integrase inhibitory peptides.

Antiretroviral peptides are a kind of bioactive peptides that present inhibitory activity against retroviruses through various mechanisms. Among them, viral integrase inhibitory peptides (VINIPs) are a class of antiretroviral peptides that have the ability to block the action of integrase proteins, which is essential for retroviral replication. As the number of experimentally verified bioactive peptides has increased significantly, the lack of in silico machine learning approaches can effectively predict the peptides with the integrase inhibitory activity. Here, we have developed the first prediction model for identifying the novel VINIPs using the sequence characteristics, and the hybrid feature set was considered to improve the predictive ability. The performance was evaluated by 5-fold cross-validation based on the training dataset, and the result indicates the proposed model is capable of predicting the VINIPs, with a sensitivity of 85.82%, a specificity of 88.81%, an accuracy of 88.37%, a balanced accuracy of 87.32% and a Matthews correlation coefficient value of 0.64. Most importantly, the model also consistently provides effective performance in independent testing. To sum up, we propose the first computational approach for identifying and characterizing the VINIPs, which can be considered novel antiretroviral therapy agents. Ultimately, to facilitate further research and development, iDVIP, an automatic computational tool that predicts the VINIPs has been developed, which is now freely available at http://mer.hc.mmh.org.tw/iDVIP/.

Elucidation of Antiviral and Antioxidant Potential of C-Phycocyanin against HIV-1 Infection through In Silico and In Vitro Approaches.

Antiretroviral therapy is the single existing therapy for patients infected with HIV; however, it has drawbacks in terms of toxicity and resistance. Thus, there is a continuous need to explore safe and efficacious anti-retroviral agents. C-Phycocyanin (C-PC) is a phycobiliprotein, which has been known for various biological properties; however, its effect on HIV-1 replication needs revelation. This study aimed to identify the inhibitory effects of C-PC on HIV-1 using in vitro and in silico approaches and to assess its role in the generation of mitochondrial reactive oxygen species (ROS) during HIV-1 infection. In vitro anti-HIV-1 activity of C-PC was assessed on TZM-bl cells through luciferase gene assay against four different clades of HIV-1 strains in a dose-dependent manner. Results were confirmed in PBMCs, using the HIV-1 p24 antigen assay. Strong associations between C-PC and HIV-1 proteins were observed through in silico molecular simulation-based interactions, and the in vitro mechanistic study confirmed its target by inhibition of reverse transcriptase and protease enzymes. Additionally, the generation of mitochondrial ROS was detected by the MitoSOX and DCF-DA probe through confocal microscopy. Furthermore, our results confirmed that C-PC treatment notably subdued the fluorescence in the presence of the virus, thus reduction of ROS and the activation of caspase-3/7 in HIV-1-infected cells. Overall, our study suggests C-PC as a potent and broad in vitro antiviral and antioxidant agent against HIV-1 infection.

POS-127 SPECTRUM OF RENAL DISEASE IN PATIENTS WITH RETROVIRAL DISEASE AT TERITIARY CARE CENTER

Patterns of HIV associated renal disease varies from acute kidney injury (AKI), HIV associated nephropathy (HIVAN), immune complex mediated, chronic kidney disease (CKD), and electrolyte and acid base disorders. Prerenal causes like dehydration, gastroenteritis, pancreatitis, and intrinsic causes like infections, septic shock, and antiretroviral agents, analgesics, toxins, and metabolic syndrome associated with antiretroviral therapy use, CKD with incidental HIV infection. Intrinsic renal disease like ischemic renal injury due to sepsis is most common cause of AKI, rare causes like post renal cause of AKI in HIV like medication induced crystalluria, hyperuricosuria, pelvic lymphadenopathy, retoperitoneal fibrosis.

Animal models for studies of HIV-1 brain reservoirs.

The HIV-1 often evades a robust antiretroviral-mediated immune response, leading to persistent infection within anatomically privileged sites including the CNS. Continuous low-level infection occurs in the presence of effective antiretroviral therapy (ART) in CD4+ T cells and mononuclear phagocytes (MP; monocytes, macrophages, microglia, and dendritic cells). Within the CNS, productive viral infection is found exclusively in microglia and meningeal, perivascular, and choroidal macrophages. MPs serve as the principal viral CNS reservoir. Animal models have been developed to recapitulate natural human HIV-1 infection. These include nonhuman primates, humanized mice, EcoHIV, and transgenic rodent models. These models have been used to study disease pathobiology, antiretroviral and immune modulatory agents, viral reservoirs, and eradication strategies. However, each of these models are limited to specific component(s) of human disease. Indeed, HIV-1 species specificity must drive therapeutic and cure studies. These have been studied in several model systems reflective of latent infections, specifically in MP (myeloid, monocyte, macrophages, microglia, and histiocyte cell) populations. Therefore, additional small animal models that allow productive viral replication to enable viral carriage into the brain and the virus-susceptible MPs are needed. To this end, this review serves to outline animal models currently available to study myeloid brain reservoirs and highlight areas that are lacking and require future research to more effectively study disease-specific events that could be useful for viral eradication studies both in and outside the CNS.

A CROSS-SECTIONAL ASSESSMENT OF PHARMACISTS' KNOWLEDGE, ATTITUDE AND PRACTICE OF PREVENTION OF MOTHER-TO-CHILD TRANSMISSION OF HIV IN TWO NIGERIAN TEACHING HOSPITALS.

Background:The pharmacological component of prevention of mother-to-child transmission (PMTCT) services involves the provision of antiretroviral agents (ARVs) to the mothers and/or their babies at any stage of pregnancy. This study assessed the knowledge, attitude and practice (KAP) of Pharmacists about PMTCT.Materials and Methods:A questionnaire-based cross-sectional study was conducted among consenting Pharmacists at Ahmadu Bello University Teaching Hospital (ABUTH) and University of Nigeria Teaching Hospital (UNTH). Completed questionnaires were collated and analyzed using SPSS Version-25 with appropriate descriptive and inferential statistics. P-values less than 0.05 were considered to be statistically significant.Results:A total of 77 Pharmacists participated in the study, with 54(70.13%) being from ABUTH. In ABUTH, 15(33.3%) Pharmacists identified as being females, against 16(69.6%) in UNTH. Majority (40,95.2%) of the Pharmacists in ABUTH had less than 10 years working experience as against 8(34.8%) in UNTH. Forty-eight (88.9%) respondents knew the correct meaning of PMTCT. The Pharmacists in ABUTH and UNTH had mean knowledge scores of 58.70±2.88% and 52.17±6.19%, respectively; t(75)=1.094, p=0.760. In ABUTH and UNTH, 16(69.6%) and 22(42.3%) Pharmacists, respectively, strongly agreed that PMTCT can prevent future infections in the infants. Their mean attitude scores were 69.65±1.22% (ABUTH) and 74.09±1.68% (UNTH); t(73)=-2.063, p=0.487. For practice, 4(5.33%) Pharmacists in both hospitals very often dispensed PMTCT drugs, while 37(70.83%) counseled PMTCT treatment-naïve patients.Conclusion:The Pharmacists assessed in both hospitals had a fair knowledge of PMTCT services. Their attitudes to PMTCT was very good, although only a few of them had experience in providing care for PMTCT patients.

A CROSS-SECTIONAL ASSESSMENT OF PHARMACISTS’ KNOWLEDGE, ATTITUDE AND PRACTICE OF PREVENTION OF MOTHER-TO-CHILD TRANSMISSION OF HIV IN TWO NIGERIAN TEACHING HOSPITALS

Background: The pharmacological component of prevention of mother-to-child transmission (PMTCT) services involves the provision of antiretroviral agents (ARVs) to the mothers and/or their babies at any stage of pregnancy. This study assessed the knowledge, attitude and practice (KAP) of Pharmacists about PMTCT. Materials and Methods: A questionnaire-based cross-sectional study was conducted among consenting Pharmacists at Ahmadu Bello University Teaching Hospital (ABUTH) and University of Nigeria Teaching Hospital (UNTH). Completed questionnaires were collated and analyzed using SPSS Version-25 with appropriate descriptive and inferential statistics. P-values less than 0.05 were considered to be statistically significant. Results: A total of 77 Pharmacists participated in the study, with 54(70.13%) being from ABUTH. In ABUTH, 15(33.3%) Pharmacists identified as being females, against 16(69.6%) in UNTH. Majority (40,95.2%) of the Pharmacists in ABUTH had less than 10 years working experience as against 8(34.8%) in UNTH. Forty-eight (88.9%) respondents knew the correct meaning of PMTCT. The Pharmacists in ABUTH and UNTH had mean knowledge scores of 58.70±2.88% and 52.17±6.19%, respectively; t(75)=1.094, p=0.760. In ABUTH and UNTH, 16(69.6%) and 22(42.3%) Pharmacists, respectively, strongly agreed that PMTCT can prevent future infections in the infants. Their mean attitude scores were 69.65±1.22% (ABUTH) and 74.09±1.68% (UNTH); t(73)=-2.063, p=0.487. For practice, 4(5.33%) Pharmacists in both hospitals very often dispensed PMTCT drugs, while 37(70.83%) counseled PMTCT treatment-naïve patients. Conclusion: The Pharmacists assessed in both hospitals had a fair knowledge of PMTCT services. Their attitudes to PMTCT was very good, although only a few of them had experience in providing care for PMTCT patients.

Overview of Forced Degradation Analysis for FDA Approved Antiretroviral agents: A Review

Published on:August 2022Journal of Young Pharmacists, 2022; 14(3):273-282Review Article | doi:10.5530/jyp.2022.14.55 Authors:Suman Lamichhane1, Bhaswati Das2, Rudra Prasad Adhikari1, Mari Raju Jeyaprakash1*1Department of Pharmaceutical Analysis, JSS College of Pharmacy, JSS Academy of Higher Education and Research, Ooty, Nilgiris, Tamil Nadu, INDIA.2Department of Pharmaceutics, JSS College of Pharmacy, JSS Academy of Higher Education and Research, Ooty, Nilgiris, Tamil Nadu, INDIA. Abstract: Forced degradation and its analysis play a crucial role in the development and production of pharmaceutical products at various stages. Such studies include the assessment of appropriate drug candidates, product development, as well as comparability studies, clarification of probable intermediates and identification of by-products, and the development of stability-indicating technologies. The stability of drugs can be analyzed with Forced degradation study, which gives perspective knowledge of the molecule’s stability as well as the degradant that is produced during the drug’s shelf life. In this review on stability-indicating methods for FDA approved antiretroviral drugs from (1987-2021) such as Zidovudine, Lamivudine, Nevirapine, Ritonavir, Abacavir, Efavirenz, Tenofovir, Atazanavir, Emtricitabine, Enfuvirtide, Fosamprenavir, Tipranavir, Darunavir, Maraviroc, Raltegravir Etravirine, Nevirapine, Rilpivirine, Dolutegravir, Cobistat, Doravirine, Fostemsavir, and Cabotegravir is mentioned and analytical tools like HPLC, LC-MS/MS, UPLC their different parameters for the chromatographic condition are mentioned. No such guideline is available that mentions the pH for hydrolysis, the temperature at which thermal deterioration occurs, and the concentration of oxidation agent for oxidative degradation. The regulatory necessities for the shortcomings of the above-mentioned methods were highlighted. As a result, this study addresses recent approaches for forced degradation by presenting techniques for accessing studies on degradation mechanisms, this review helps researchers to get information on stability-indicating methods for FDA Approved antiretroviral agents for their development and validation of stability indicating methods and the characterization of degradation products produced during degradation studies. Keywords: Stability indicating methods, Anti-retroviral, Chromatographic technique, HPLC, LC-MS/MS, UPLC.

The antiretroviral agents azidothymidine, stavudine, and didanosine have the identical mutational fingerprint in the rpoB region of Escherichia coli.

We looked at the mutational fingerprints of three antiretroviral (anti-HIV) agents, azidothymidine (AZT), stavudine (STAV), and didanosine (DIDA) in the rpoB system of Escherichia coli and compared them with each other and with the fingerprints of trimethoprim and of spontaneous mutations in a wild-type and a mutT background. All three agents gave virtually identical fingerprints in the wild-type background, causing only A:T→C:G changes at 3 of the 12 A:T→C:G possible sites among the total of 92 possible base substitution mutations, even though AZT and STAV are thymidine analogs but DIDA is an adenosine analog. As all three agents are reverse transcriptase inhibitors, and act as chain blockers, the common fingerprint may be a property of chain blocking agents.

Development of Synthetic Approaches Towards HIV Integrase Strand Transfer Inhibitors (INSTIs)

AbstractA clear and precise report on synthetic methods for INSTIs and their intermediates are provided in the present work. HIV Integrase strand transfer inhibitors (INSTIs), a class of antiretroviral agents employed to treat HIV infected patients are placed as the second option of treatment after nucleosides. As INSTIs are employed as first line therapy to newly infected HIV positive individuals, thus synthetic routes must be easily accessible to the medicinal chemists towards lead optimization or process development. The current perspective elaborates synthetic routes of first to next generation INSTIs investigated in two decades of research and their development efforts. Further, drug resistance occurred by mutations against usage of INSTI drugs with respect to their generation also discussed. Later, discussion about synthetic development and investigation of next generation INSTIs against drug resistance HIV infections also covers to fill the gap between first/second generation INSTIs. The current perspective will draw attention of medicinal chemists and multi‐disciplinary researchers in HIV drug discovery.

Pharmacists and the next generation of HIV pre-exposure prophylaxis

The use of pre-exposure prophylaxis (PrEP) has curtailed transmission of human immunodeficiency virus (HIV) in most industrialized nations. Despite the availability of PrEP, HIV infections continue to occur. Access and adherence are major limitations to PrEP. Long-acting injectable antiretroviral agents are now being added to the HIV armamentarium. Long-acting agents offer the advantage of less frequent dosing with intramuscular administration versus oral dosing. Cost, injection site pain, and needle hesitancy may be detriments to the use of these agents in certain patients. More studies are needed to completely discern the role of various products and future long-acting agents as options for PrEP. Even when we consider their limitations, long-acting agents may provide an eventual opportunity to transform the delivery of PrEP. Pharmacists should be aware of the potential impact of PrEP and the advantages and disadvantages of these new long-acting agents. In addition, pharmacists should proactively advocate for and contribute to PrEP expansion efforts.

Dolutegravir Inhibits Proliferation and Motility of BT-20 Tumor Cells Through Inhibition of Human Endogenous Retrovirus Type K.

Increasing evidence points to the role of endogenous retroviruses (ERVs) in driving cancer cell proliferation. The purpose of this study was to explore the possibility of repurposing antiretroviral agents to inhibit ERVs as a new approach to cancer treatment. We found that an integrase strand-transfer inhibitor, dolutegravir (DTG), effectively inhibited the proliferation of multiple cancer cell lines and its antiproliferative potency was positively correlated with the expression levels of the human endogenous retrovirus type K (HERV-K). DTG inhibited the expression of HERV-K in multiple human cancer cell lines and the mouse mammary tumor virus (MMTV) in the murine 4T1 mammary cancer cell line. We chose the fast-growing BT-20 cell line as a model to study the in vitro antiproliferative mechanisms of DTG. BT-20 cells overexpressing both HERV-K env and pol genes became more resistant to DTG than cells transduced with vector alone. Knockdown of HERV-K also increased DTG resistance of BT-20 cells. The antiproliferative effect of DTG correlated with enhanced expression of E-cadherin and reduction in cell motility and invasiveness. Surprisingly, DTG stimulated expression of the env gene of MMTV in vivo and promoted metastasis of 4T1 tumor cells to the lungs. Taken together, our data support the role of ERVs in tumor development and encourage the further search for antiretroviral agents to treat malignancies in which ERVs are active.

Approaches to accelerating the study of new antiretrovirals in pregnancy.

IntroductionWomen who are pregnant or who could become pregnant experience delayed access to or underinformed use of important new antiretroviral (ARV) drugs because of traditional drug development processes that ostensibly aim to reduce potential harm but effectively fail to ensure that timely information about safe and effective use in pregnancy is available.DiscussionThe World Health Organization and International Maternal, Pediatric, Adolescent Antiretroviral Clinical Trials Network convened a year‐long workshop on “Approaches to Enhance and Accelerate Study of New Drugs for HIV and Associated Infections in Pregnant Women.” Workshop participants were tasked with defining key principles and optimal approaches to including pregnant women in pre‐ and post‐licensure trials in order to accelerate the availability of pharmacokinetic and safety data for new ARV agents in pregnancy. ARV efficacy in pregnancy and ARV efficacy for prevention of vertical transmission can be extrapolated from proof of efficacy in non‐pregnant adults, provided that drug levels in pregnancy are similar. However, short‐term safety and pharmacokinetics must be studied directly in pregnant women and should be conducted and included in initial licensure for all new ARVs. Accelerating the timeline for completion of pre‐clinical studies is essential for pregnancy short‐term safety and pharmacokinetic studies to be safely completed by the time a drug is licensed. Composite key pregnancy, birth and neonatal outcomes are critical for drugs expected to have broad use, and studies should be initiated at or soon after drug licensure. Teratogenicity risk cannot be feasibly assessed before drug licensure and will depend on robust post‐marketing surveillance systems. With some modifications, these principles will apply to ARVs used for prevention, two‐drug regimens, long‐acting ARVs and ARVs administered through novel delivery systems.ConclusionsImplementation of the proposed principles and framework will enhance and accelerate the study of new ARVs in pregnancy, resulting in more timely, equitable and informed access to new ARVs for pregnant women.