A comprehensive chemical investigation of the extract from Phyllanthus hainanensis led to the isolation and identification of four new aromatic diterpenoids, phainanones A-D (1-4), along with one known analogue (5). Phainanones A (1) and B (2) are novel ent-cleistanthane-type diterpenoids, with 1 characterized as a 17-nor derivative and 2 as a 3-nor derivative, both sharing a rearranged 5/6/6 tricyclic ring system. Their structures with absolute configurations were established by comprehensive spectroscopic analysis, ECD calculations, and X-ray crystallography. Putative biosynthetic pathways for compounds 1 and 2 were also proposed. In biological assays, compounds 2-4 potently inhibited the HIV-1 Bal pseudovirus while showing low cytotoxicity (IC50 0.54-3.3 μM; CC50 > 30). The most active compound 2 also exhibited strong activity against HIV-1IIIB (IC50 = 1.0 ± 0.11 μM, SI > 98). Furthermore, its time-dependent inhibition profile indicates that it may act at a postentry step, interfering with early viral replication events such as reverse transcription. This study highlights compound 2 as a promising lead for anti-HIV drug development.

CYP46A1 activation by low-dose efavirenz uncovers the link between brain cholesterol metabolism, energetics, and vasculature.

Dihydropyrimidinones (DHPMs), also referred to as pyrimidinones, are privileged structures widely employed in the search for new compounds capable of inhibiting HIV replication. This core is found in natural alkaloids isolated from marine sponge species, which are known to inhibit the binding of the viral surface protein gp120 to the CD4 receptor of target cells. In addition, DHPMs exhibit a close structural relationship with nucleic acids, macromolecules that constitute the genetic material of both living organisms and viruses. Furthermore, the DHPM nucleus is present in several important anti-HIV drugs that inhibit reverse transcriptase and integrase enzymes, indicating that the use of these privileged structures represents a faster and more promising approach for the development of novel HIV inhibitors. In this review, we provide an overview of DHPM-based compounds as potential future trends in AIDS treatment, with a focus on studies published over the last 10 years. Overall, our analysis indicates that compounds containing the DHPM core generally display high potency against multiple HIV targets and may overcome antiviral resistance, reinforcing the relevance of this pharmacophoric fragment for the development of effective and innovative antiretroviral therapies.

BackgroundChronic kidney disease (CKD) is a frequent non-communicable complication in people living with HIV (PLHIV), influenced by antiretroviral therapy (ART), comorbidities, and healthcare setting. The Data Collection on Adverse Events of Anti-HIV Drugs (DAD) CKD risk score is widely applied internationally, while the Thai CKD risk score, developed for the general Thai population, has not been validated in HIV cohorts. This study compared the predictive accuracy of the DAD models and the Thai CKD risk score in Thai people living with HIV receiving ART.MethodsA retrospective cohort analysis was conducted using electronic medical records of adults (≥18years) living with HIV at Warinchamrap Hospital between January 2020 and May 2024. Patients with pre-existing CKD or incomplete data were excluded. CKD was defined per Kidney Disease: Improving Global Outcomes (KDIGO) criteria. Model calibration was assessed with the Hosmer-Lemeshow test, and discrimination with the C-statistic.ResultsAmong 901 PLHIV receiving ART, 104 (11.5%) developed incident CKD during a median follow-up of approximately 4years. Patients who developed CKD were more likely to be male and have a lower baseline estimated glomerular filtration rate, and higher prevalence of proteinuria and comorbidities. All evaluated models stratified CKD risk; however, the full and short DAD scores showed moderate discrimination (C-statistics ≈0.76) with evidence of risk underestimation and suboptimal calibration. The Thai CKD risk score demonstrated higher discrimination (C-statistic 0.93), sensitivity (88.5%), and better calibration in this cohort.ConclusionsIn this Thai HIV cohort, all evaluated risk models provided CKD risk stratification, but the Thai CKD risk score showed more reliable discrimination and calibration than the DAD models. These findings highlight the importance of population-specific validation when applying CKD risk prediction tools in HIV care.

Covariate adjustment in randomized clinical trials improves statistical precision and power compared to unadjusted analyses. A simple covariate adjustment method is direct regression adjustment, which uses the estimated treatment coefficient in a regression model as the effect estimate (e.g., analysis of covariance for continuous outcomes). Although maximum likelihood estimation does not yield model-robust direct regression adjustment for binary outcomes, such adjustment can be obtained via working normal linear regression for risk difference and working Poisson regression for risk ratio. This article examines the theoretical and empirical efficiency of these methods under misspecified regression models. We show that, asymptotically, they are as efficient as or more efficient than unadjusted analyses, strictly for working normal linear regression and typically for working Poisson regression. Efficiency gain is explained by reduced residual variation due to covariate adjustment. A simulation study showed that, even under model misspecification, these methods achieve efficiency comparable to logistic regression standardization with correctly specified models in a usual range of the outcome–covariate associations. Analysis of a randomized anti-HIV drug trial corroborated these findings. Direct regression adjustment using working normal linear and working Poisson regression provides simple, robust, and efficient methods for covariate adjustment in randomized trials with binary outcomes.

Although numerous, effective antiviral therapies are in clinical use, there is a significant demand for novel, improved drug delivery systems (DDS) to enhance the biological and pharmacokinetic properties of administered drugs. Nanostructured carrier systems are increasingly recognized as promising candidates; however, their development is still in its infancy. Herein, we have developed a biocompatible system composed of polylactide and chitosan, loaded with tenofovir alafenamide (TAF) as an antiretroviral drug. Our nanoparticles (NPs) were able to release TAF for 21days, depending on the chitosan content in their matrix. Moreover, the NPs were not cytotoxic and showed high antiviral activity in an in vitro HIV infection assay. Most importantly, the effectiveness of the selected nanoformulations was comparable to that of free TAF (IC50 of 56nM versus 62-75nM for NPs), indicating that TAF encapsulation preserved its antiviral effect. The results of this study demonstrate the potential of TAF-loaded NPs and provide a straightforward, effective, and biocompatible strategy for the delivery of anti-HIV drugs.

BackgroundPeople living with HIV (PLWH) exhibit two-fold higher incidence of cardiovascular disease compared to HIV-negative persons. However, predictors of cardiovascular disease risk in PLWH are still evolving. The objective of this study is to evaluate the predictors of cardiovascular disease among PLWH in Nigeria.MethodsThis cross-sectional study was conducted among adult patients attending a large HIV clinic in Kano, northern Nigeria. We used systematic sampling to recruit participants and computed their 5-year projected CVD risk using the Data collection on Adverse effects of Anti-HIV Drugs (DAD) equation.ResultsThe majority of participants were female (70.6%). The estimated median 5-year CVD risk was 0.7% (interquartile range, IQR 0.4, 10). The majority of participants (58.9%) had a low risk of developing cardiovascular disease, while 28.9% had a moderate risk. Cardiovascular disease was associated with elevated high-sensitivity C-reactive protein (hsCRP) > 3.03 mg/L [adjusted odds ratio, aOR: 4.58, 95% CI: 2.09–10.04), p = 0.001], increasing age [aOR 2.38, 95% CI (1.48–4.50), p = 0.020], male sex [aOR 2.16, 95% CI (1.03–4.53), p = 0.040] and hypercholesterolemia [aOR 3.03, 95% CI (1.68–4.86), p = 0.005].ConclusionThe majority of PLWH in our setting have low to moderate risk of developing cardiovascular disease. Cardiovascular disease risk was associated with elevated hsCRP, increasing age, male sex, and hypercholesterolemia. Our findings highlight the importance of early CVD risk stratification to prevent morbidity and mortality among PLWH.

Agreement and Fairness of Cardiovascular Risk Assessment Models Among Older People Living With HIV in China: A Multicenter Cross-sectional Study.

People living with HIV (PLHIV) face greater cardiovascular risk but lack targeted prevention, especially in Asia. This study compares cardiovascular risk models among PLHIV in Malaysia. We included PLHIV aged ≥18 years between January 2016 and December 2017. Those with pre-existing cardiovascular disease (CVD) or non-Malaysians were excluded. Events were first cardiovascular hospitalization or death within five years. Four models were assessed: Framingham risk score (FRS), Revised Pooled Cohort Equations (RPCE), Data Collection on Adverse Effects of Anti-HIV Drugs (D:A:D) full and reduced. Discrimination was assessed with c-statistics and calibration with observed-to-expected (O/E) ratios. Among 6,339 PLHIV, 136 (2.1%) developed CVD. Those with CVD more often had dyslipidemia (22.8% vs. 16.8%), hypertension (23.5% vs. 7.2%), and diabetes (15.4% vs. 4.2%). All models showed similar moderate discrimination (c-statistics: 0.75-0.77, 95% CI: 0.71-0.81). D:A:D reduced calibrated better (O/E: 1.13, 95% CI: 0.94-1.32) than the full version (1.25, 95% CI: 1.04-1.46). Framingham risk score (FRS) (0.58, 95% CI: 0.49-0.68) and RPCE (0.72, 95% CI: 0.58-0.90) overestimated risk. While all models showed similar discrimination, D:A:D reduced provided the most accurate 5-year estimates with greater practicality. This supports its use for scalable cardiovascular risk assessment in HIV care.

Regression-based analysis of topological indices in simple molecular graphs of anti-HIV drug compounds

Objective: The goal of this study was to use a Quality by Design (QbD) approach to create and improve liposomal formulations of Emtricitabine, an anti-HIV drug, to make it better at trapping other substances. Methods: A Box-Behnken Design (BBD) with three factors and three levels was used to systematically look at how the independent variables- Soy Lecithin (60–70%), Cholesterol (20–30%), and Palmitic Acid (5–10%)—affected the dependent variable, Entrapment Efficiency (EE %). We made and tested fifteen different formulas. Results: The statistical analysis of the BBD indicated that a Linear model was sufficient to describe the design space, as higher-order models were not significant. The ANOVA for the linear model yielded an F-value of 1.00 (p=0.4289), signifying that the model was not significant in relation to noise. None of the individual factors (Soy Lecithin, p=0.5831; Cholesterol, p=0.2393; Palmitic Acid, p=0.3099) demonstrated a statistically significant effect on EE within the studied ranges. However, the lack of fit was non-significant (p=0.9515), confirming the model's adequacy. A confirmation batch prepared with 70% Soy Lecithin, 20% Cholesterol, and 10% Palmitic Acid yielded an EE of 66.96%, which was within the 95% prediction interval (53.21% - 81.30%) of the predicted value (67.25%). Conclusion: Liposomal formulation of Emtricitabine was optimised and prepared with significant entrapment efficiency, which made possible to test it further through in vitro and in vivo studies. Keywords: Emtricitabine, Liposomes, Box-Behnken Design, Entrapment Efficiency, Quality by Design, Optimization, Anti-HIV.

The persistence of human immunodeficiency virus (HIV) reservoirs in the lymphatic system remains a major obstacle to a complete cure, largely due to subtherapeutic concentrations of antiretrovirals (ARVs) in lymphatic tissues under current regimens. Developing innovative strategies to enhance ARV delivery to lymphatic tissues could therefore represent a paradigm shift in the pursuit of an HIV cure. Here, we describe a delivery strategy involving an Evans Blue (EB)-based prodrug conjugated with dolutegravir (DTG), a model ARV. This construct is designed to bind albumin with high affinity, facilitating transport to lymph nodes (LNs) via albumin-mediated trans-endothelial transcytosis. EB modification substantially improved the distribution of the conjugated molecule to LNs throughout the body, and the hEB-DTG prodrug increased DTG concentrations in LNs by up to 10-fold, suggesting a promising strategy for eradicating HIV reservoirs. STATEMENT OF SIGNIFICANCE: A major barrier to curing HIV infection is the persistence of viral reservoirs in lymphatic tissues, which current ARV therapies fail to eliminate due to poor drug penetration. To overcome this limitation, we developed a prodrug strategy that harnesses the natural trafficking of albumin to the lymphatic system. In this approach, ARVs are conjugated to EB, a high-affinity albumin-binding dye, to generate EB-ARV prodrugs that hijack albumin's endogenous transport pathways. After intravenous administration, these conjugates rapidly associate with circulating albumin, which mediates their transcytosis across endothelial barriers and directs their accumulation in lymph nodes and other lymphoid tissues. This strategy substantially enhances ARV exposure at the primary sites of viral persistence, representing a promising and potentially transformative approach for eradicating HIV reservoirs and advancing toward a functional cure.

The development of new anti-HIV drugs with a novel mechanism of action is still required. A human cell surface protein, CD4, is a primary receptor for HIV-1 entry, and we have developed small-molecule CD4-competitive HIV-1 entry inhibitors, including CD4 mimics. CD4 mimics bind to a viral envelope protein, gp120, and cause a conformational change. Combinational use of CD4 mimics with CD4-induced antibodies, which recognize the exposed regions of gp120 induced by the CD4/CD4 mimic binding, shows a synergistic effect of anti-HIV activity. Therefore, we envisioned that antibody-drug conjugates (ADCs) of CD4 mimics and CD4-induced antibodies would effectively inhibit virus entry. Herein, ADCs of CD4 mimics and CD4-induced antibodies were designed and synthesized, and their anti-HIV-1 activities were evaluated. As a result, several synthesized ADCs exhibited 7- to 10-fold higher anti-HIV-1 activity than that of the parent antibodies, although there remains significant room for improvement in these ADCs.

In this study, a quantitative structure-property relationship (QSPR) analysis was conducted to predict the physiochemical properties of anti-HIV drug molecules using a linear regression model. The model utilized neighborhood degree sum topological indices, which are graph-theoretical descriptors representing molecular structure, as key predictive features. These indices were calculated for each molecule, providing a numerical representation of their structural properties. The linear regression model effectively correlated these indices with the known physicochemical properties of the drugs, demonstrating its potential to predict the efficacy of new compounds. This approach offers a valuable tool for designing and optimizing anti-HIV drugs based on molecular topological descriptors.

Globally, a significant overlap exists between females affected by HIV and HSV-2 infections, and who have an unmet need for contraception. Intravaginal rings (IVRs) have become widely accepted by women worldwide for contraception and hormone replacement therapy and provide a promising platform as a multipurpose prevention technology (MPT). Using state-of-the-art 3D printing process known as continuous liquid interface production (CLIP™), IVRs with an internal honeycomb (HC 2.53mm) geometry were fabricated with a silicone-urethane resin. IVRs were loaded with a triple-drug combination of an anti-HIV drug (Dapivirine, DPV, 30mg), an anti-herpes drug (Pritelivir, PTV, 20mg) and a contraceptive hormone (Levonorgestrel, LNG, 2.0mg) using a solvent swelling method in acetone. IVRs elicited zero-order release kinetics following an initial burst for all three APIs when formulated individually or in combination. Release rates were above benchmark therapeutic targets for DPV and LNG (200µg/day DPV, 20µg/day LNG). A series of accelerated stability studies demonstrated the physical integrity of IVRs after 6months of storage at 40ºC/75%RH. DPV remained stable over 6months, whereas PTV and LNG exhibited significant decrease in concentration after 3months of storage with presence of degradation products detected by HPLC. Mouse size placebo rings (3mm OD) elicited 100% cell viability in relevant cell lines and were well tolerated in vivo in mice. Collectively, these results demonstrate that this first-in-line 3D printed MPT IVR has potential to expand preventative choices for young women and girls against HIV, HSV, and unplanned pregnancy.

Ultrasensitive detection of anti-HIV drug nevirapine in nanomolar level based on nickel‑copper bimetallic MOF composites sensing platform

Reverse transcriptase (RT), as an essential key enzyme in the replication process of the human immunodeficiency virus (HIV), serves as a crucial target for the development of anti-HIV drugs. Nevertheless, the frequent mutation of RT leads to drug resistance. Therefore, there is an urgent need for the rapid identification of the drug resistant-susceptible relationship. In this study, we developed the non-nucleoside RT inhibitor (NNRTI) and RT resistant-susceptible interaction prediction model (NRIP). By introducing a descriptor incorporating sequence description of RT mutation and nonuniform spatial shell structures combined with residue properties, NRIP was trained based on 4324 pairs of NNRTIs and RT interactions through an extreme gradient boosting (XGBoost) classifier. Results of 10-fold cross-validation indicated that the baseline of the sequence-descriptor-based model could reach the ROC-AUC of 0.886, which could further be increased to 0.967 by incorporating spatial descriptors. More importantly, NRIP could achieve the ROC-AUC of 0.971 and the PR-AUC of 0.974 on the independent testing dataset. Finally, a multistep virtual screening pipeline was constructed by incorporating the NRIP model with structure similarity calculation, drug likeness assessment, and molecular docking, illustrating the potential of screening bioactive compounds of natural compounds from FOODB.

With the use of newer and more robust antiretrovirals, the risk of viral transmission through breastfeeding has sharply diminished. This has led to a change in guideline recommendations. Currently, breastfeeding is regarded as an equipoise of formula, in case of a well-controlled HIV infection. However, it is not yet fully established whether infant exposure to antiretroviral drugs through breastmilk is a reason for concern. Only sparse and heterogenous data exist on concentrations of dolutegravir, raltegravir, bictegravir and doravirine in breastmilk. So further research is required and to accurately describe infant exposure of antiretrovirals through breastmilk, reproducible bioanalytical methods are needed. Our existing UPLC-MS/MS method for several anti-HIV drugs in EDTA-plasma was modified for validation of dolutegravir, bictegravir, raltegravir and doravirine concentrations in human breastmilk. [13C,2H5]-Dolutegravir, [13C,2H2,15N]-bictegravir, [2H6]-raltegravir and [13C6]-doravirine were used as internal standard. The sample preparation involved protein precipitation and detection was performed with tandem mass spectrometry (MS/MS) in a total runtime of 10min. The assay was validated over the concentration range of 0.0100-10.0mg/L for dolutegravir and doravirine, 0.00500-10.0mg/L for raltegravir and 0.0200-20.0mg/L for bictegravir. Within-run and between-run accuracy were within ±10% of the nominal concentration and precision <10 CV% for quality controls at high, medium and low concentrations, and±8.8% and<14 CV%, respectively, at the lower limit of quantification for all analytes. Extraction recovery was 81% and 79% for dolutegravir and its internal standard, 83% and 83% for bictegravir and its internal standard, 89% and 92% for raltegravir and its internal standard and 104% and 101% for doravirine and its internal standard. Processed samples of dolutegravir, bictegravir, raltegravir and doravirine in breastmilk were 108%, 95.5%, 105% and 102% after 14days at 4°C. The assay is currently being implemented successfully in pharmacokinetic studies.

Physiologically based pharmacokinetic (PBPK) modeling is a computational technique that uses the physicochemical properties of drugs and physiological information to simulate plasma and tissue concentrations. PBPK modeling has become a mainstream approach in drug research and development, frequently employed to support regulatory packages for new drug applications. Understanding the pharmacokinetic characteristics of anti-HIV drugs is essential for successful treatment. In recent decades, PBPK modeling has been commonly used in the development and clinical therapy of anti-HIV medications. This review discusses the prevalence and application of PBPK modeling in the pharmacokinetics of anti-HIV drugs. Among the articles retrieved for this review, PBPK modeling was predominantly employed for anti-HIV drugs in contexts, such as pregnancy, drug-drug interactions, and pediatrics. The most commonly used software programs for this model are Simcyp, MATLAB, and PK-sim. This review will provide insights for researchers in applying PBPK models to manage patients with HIV infection, aiming to enhance the efficacy of anti-HIV drug therapy and prevent undesirable adverse effects.

The current analytical work focuses on the development of a bioanalytical method for anti-HIV drugs, nevirapine, fosamprenavir calcium, and its metabolite amprenavir. Fosamprenavir is a prodrug of amprenavir, digested by cellular phosphatases in vivo. Rat plasma was isolated from the collected rat blood by centrifugation and used for preparing stock solutions. A reverse-phase column (ODS C-18) was used for the development of the method and validation. The bioanalytical method was developed using the isocratic mode. The solvent system used is Methanol: Water: ACN (800mL: 200mL: 50mL). The method was run at a flow rate of 0.5mL/min, and 258nm was the detection wavelength. The developed method showed retention at 8.705min for nevirapine, 11.923min for fosamprenavir calcium, and 14.391min for amprenavir. The calibration curve was linear with a correlation coefficient (r2) of 0.9916 for nevirapine, 0.9909 for fosamprenavir calcium, and 0.9879min for amprenavir. The RSD of the accuracy, precision, and stability study of the method was found to be less than 2% and was found to be acceptable. The method is reliable and does not show any kind of interference due to the plasma sample. Thus, the results support that a reliable, reproducible, and efficient method was developed, and validation was carried out for the estimation of the drug nevirapine, fosamprenavir, and its metabolite amprenavir in rat plasma samples.