Nelfinavir Research Articles

Regulatory aspects of Impurity profiling

Quality, safety and efficacy of pharmaceuticals play an important role in drug therapy. The safety attribute of drug is established by its pharmacological or toxicological profile along with adverse effects caused by impurities in bulk and dosage form. Impurities present in drug often possess undesired pharmacological or toxicological effects which outweighs the benefits of drug therapy. Recently, many impurity cases have been reported for e.g. NDMA (N-nitroso dimethylamine) impurity in drug product Ranitidine. This may be due to inappropriate follow of impurity related regulatory guidelines or critical voids in regulatory control of impurities or may be lack of appropriate analytical technique for impurity detection. Moreover, in June 2007, EMA became aware of contamination of Viracept® tablets. It was detected with presence of ethyl mesylate impurity, a known genotoxic substance in Nelfinavir mesylate (Viracept®). Due to this, the product was recalled from EU market.
 An impure drug is devoid of safety, quality and efficacy and can lead to adverse events. So, drugs of required quality standards should reach the market for patient’s safety. Therefore it is important and becomes mandatory to submit impurity data related to isolation, identification, qualification and control of impurities to respective regulatory authorities. This article is an attempt to deliver comprehensive understanding related to various attributes and details about impurity profiling in context with regulatory guidelines along with detail description of impurity data submission to regulatory authority for drug substance (API) to get market approval. This article also focus on ICH impurity guidelines ,sources, classification and quality control of impurities.

Nelfinavir inhibits human DDI2 and potentiates cytotoxicity of proteasome inhibitors

Proteasome inhibitors (PIs) are currently used in the clinic to treat cancers such as multiple myeloma (MM). However, cancer cells often rapidly develop drug resistance towards PIs due to a compensatory mechanism mediated by nuclear factor erythroid 2 like 1 (NFE2L1) and aspartic protease DNA damage inducible 1 homolog 2 (DDI2). Following DDI2-mediated cleavage, NFE2L1 is able to induce transcription of virtually all proteasome subunit genes. Under normal condition, cleaved NFE2L1 is constantly degraded by proteasome, whereas in the presence of PIs, it accumulates and induces proteasome synthesis which in turn promotes the development of drug resistance towards PIs. Here, we report that Nelfinavir (NFV), an HIV protease inhibitor, can inhibit DDI2 activity directly. Inhibition of DDI2 by NFV effectively blocks NFE2L1 proteolysis and potentiates cytotoxicity of PIs in cancer cells. Recent clinical evidence indicated that NFV can effectively delay the refractory period of MM patients treated with PI-based therapy. Our finding hence provides a specific molecular mechanism for combinatorial therapy using NFV and PIs for treating MM and probably additional cancers.

Cover Image, Volume 92, Number 10, October 2020

Front Cover Caption: The cover image is based on the Research Article The anti-HIV drug nelfi navir mesylate (Viracept) is a potent inhibitor of cell fusion caused by the SARSCoV-2 spike (S) glycoprotein warranting further evaluation as an antiviral against COVID-19 infections, https://doi.org/10.1002/jmv.25985.

The FDA-Approved Drug Nelfinavir Inhibits Lytic Cell-Free but Not Cell-Associated Nonlytic Transmission of Human Adenovirus.

Adenoviruses (AdVs) are prevalent and give rise to chronic and recurrent disease. Human AdV (HAdV) species B and C, such as HAdV-C2, -C5, and -B14, cause respiratory disease and constitute a health threat for immunocompromised individuals. HAdV-Cs are well known for lysing cells owing to the E3 CR1-β-encoded adenovirus death protein (ADP). We previously reported a high-throughput image-based screening framework and identified an inhibitor of HAdV-C2 multiround infection, nelfinavir mesylate. Nelfinavir is the active ingredient of Viracept, an FDA-approved inhibitor of human immunodeficiency virus (HIV) aspartyl protease that is used to treat AIDS. It is not effective against single-round HAdV infections. Here, we show that nelfinavir inhibits lytic cell-free transmission of HAdV, indicated by the suppression of comet-shaped infection foci in cell culture. Comet-shaped foci occur upon convection-based transmission of cell-free viral particles from an infected cell to neighboring uninfected cells. HAdV lacking ADP was insensitive to nelfinavir but gave rise to comet-shaped foci, indicating that ADP enhances but is not required for cell lysis. This was supported by the notion that HAdV-B14 and -B14p1 lacking ADP were highly sensitive to nelfinavir, although HAdV-A31, -B3, -B7, -B11, -B16, -B21, -D8, -D30, and -D37 were less sensitive. Conspicuously, nelfinavir uncovered slow-growing round HAdV-C2 foci, independent of neutralizing antibodies in the medium, indicative of nonlytic cell-to-cell transmission. Our study demonstrates the repurposing potential of nelfinavir with postexposure efficacy against different HAdVs and describes an alternative nonlytic cell-to-cell transmission mode of HAdV.

Mutações de resistência a antirretrovirais e diversidade genética do HIV-1 em gestantes atendidas em uma maternidade de referência no Estado do Piauí, Brasil

Objetivo: identificar os subtipos do HIV-1, avaliar a presença de mutações de resistência aos antirretrovirais (DRM) e estimar o nível de resistência das cepas mutantes detectadas em mulheres gestantes atendidas na maternidade de referência no Piauí, nordeste do Brasil. Metodologia: Das 873 mulheres gestantes, 17 infectadas pelo HIV-1 foram selecionadas para participar deste estudo. A genotipagem do HIV-1 foi realizada usando sequenciamento de nucleotídeos seguida de análises filogenéticas para determinar o subtipo viral, incluindo a identificação usando ferramenta on-line COMET. DRM foram definidas usando o programa Calibrated Population Resistance Tool. Análises adicionais também foram realizadas para verificar a resistência aos medicamentos no pré-tratamento usando o Programa Stanford HIVdb. Resultados: O subtipo B foi detectado em todas as gestantes infectadas pelo HIV-1. A prevalência de DRM foi 11.8%. As mutações L90M e K103N foram identificadas, sendo estimada resistência alta (nelfinavir e nevirapine) e intermediária (indinavir e saquinavir) a diferentes antirretrovirais. Conclusão: Este estudo identificou importantes informações genéticas do HIV-1 em gestantes no Piauí, as quais podem auxiliar no direcionamento de medidas de controle e de prevenção dessa infecção viral.

A high-content image-based drug screen of clinical compounds against cell transmission of adenovirus

Human adenoviruses (HAdVs) are fatal to immuno-suppressed individuals, but no effective anti-HAdV therapy is available. Here, we present a novel image-based high-throughput screening (HTS) platform, which scores the full viral replication cycle from virus entry to dissemination of progeny and second-round infections. We analysed 1,280 small molecular weight compounds of the Prestwick Chemical Library (PCL) for interference with HAdV-C2 infection in a quadruplicate, blinded format, and performed robust image analyses and hit filtering. We present the entire set of the screening data including all images, image analyses and data processing pipelines. The data are made available at the Image Data Resource (IDR, idr0081). Our screen identified Nelfinavir mesylate as an inhibitor of HAdV-C2 multi-round plaque formation, but not single round infection. Nelfinavir has been FDA-approved for anti-retroviral therapy in humans. Our results underscore the power of image-based full cycle infection assays in identifying viral inhibitors with clinical potential.

Abstract A74: The HIV protease inhibitor nelfinavir, alone or in combination with the proteasome inhibitor bortezomib, is cytotoxic to high-grade serous ovarian cancer cells regardless of platinum sensitivity

Abstract High-grade serous ovarian cancer (HGSOC) is a deadly disease, affecting women worldwide. Major challenges in treating HGSOC patients involve overcoming resistance to platinum (Pt)-based therapy by developing novel treatment approaches. Nelfinavir (NFV), an anti-HIV drug, has shown to be effective against cancers of high secretory capacity—e.g., multiple myeloma—by aggravating the stress of the endoplasmic reticulum (ER). Its potential therapeutic role against HGSOC, however, remains unclear. We hypothesized that simultaneous ER stress aggravation by NFV and blockage of the proteolytic capacity of the proteasome with a proteasome inhibitor should cause sufficient ER stress to tilt cells to a death fate. In this study, we investigated the cytotoxicity of the ER aggravator NFV as monotherapy and in combination with the proteasome inhibitor bortezomib (BZ) against HGSOC cells having differential sensitivities to cisplatin (CDDP). NFV caused cell cycle arrest associated with increased expression of cyclin-dependent kinase inhibitor p27kip1, while triggering the unfolded protein response (UPR) in a dose-dependent manner and assessed by increased subrogate ER stress biomarkers GRP78 and CHOP. We also discovered that blocking the ubiquitin proteasome system (UPS) using cytostatic concentrations of BZ causes accumulation of poly-ubiquitinated proteins and further activation of the UPR. More importantly, when we combined BZ with NFV, we observed a potentiation of the action of BZ leading to HGSOC lethality regardless of Pt sensitivity. These results suggest that NFV could potentially be used following standard Pt-based treatment, either as monotherapy or in conjunction with proteasome inhibitors, to kill the remaining HGSOC cells that otherwise recur and recreate the disease. In addition, the data provide the rationale for rapid repurposing to treat HGSOC of two drugs clinically approved for other uses, as their systemic toxicities have already been assessed. Citation Format: Mahbuba R. Subeha, Alicia A. Goyeneche, Michael A. Lisio, Carlos M. Telleria. The HIV protease inhibitor nelfinavir, alone or in combination with the proteasome inhibitor bortezomib, is cytotoxic to high-grade serous ovarian cancer cells regardless of platinum sensitivity [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr A74.

Anti-HIV and Anti-HCV drugs are the putative inhibitors of RNA-dependent-RNA polymerase activity of NSP12 of the SARS CoV-2 (COVID-19)

Corona virus is the emerging infectious viral agent that had to threaten the world with its epidemic since 2003 with SARS and 2013 with MERS virus. This virus again stuck the whole world with its robust pandemic in 2019whena novel form of the corona virus was emerged in and infect the local population of Wuhan, China. The virus named as SARS CoV-2 and the disease caused by this infectious virus is introduced as COVID 19. Since December 2019, this virus took more than 432, 973 lives all around the world. This study elaborates the role of HIV and HCV drug in targeting nsp12 gene of SARS CoV 2 that is responsible for RNA dependent RNA polymerase activity. The work employed homology modelling, structure validation and molecular docking analysis to evaluate the binding affinity and interaction analysis between NSP12 (RdRp) protein and HIV/HCV drugs. Outcome of the study impacted on Nelfinavir (NFV), Raltegravir (RAL) and Delavirdine (DLV) among Anti-HIV and Paritaprevir (PTV), Beclabuvir (BCV) and Ledipasvir (LDV) among Anti-HCV as the most effective inhibitors of SARS CoV-2 RdRp ternary complexes. The drugs have a strong binding affinity with the residues that are present in the active site of RdRp of the virus and essential for its replication. This study establishes significant information in the direction of therapeutic development as we are dealing with the situation where we urgently required medication or vaccine to combat COVID-19. Therefore, this study provides essential molecular information about the FDA approved antiviral drugs that can be used to treat this disease. Importantly, the listed drugs had never prioritized for their effectiveness against COVID-19.

Nanotechnology Based Repositioning of an Anti-Viral Drug for Non-Small Cell Lung Cancer (NSCLC).

Nelfinavir (NFV), a FDA approved antiretroviral drug, has been reported to exhibit cancer cells growth inhibition and increased apoptosis. However, it requires a higher dose leading to toxicity, thus limiting its potential clinical translation. We aim to develop biodegradable (poly (lactic-co-glycolic acid)) PLGA nanoparticles of nelfinavir and determine their efficacy to treat non-small cell lung cancer (NSCLC). HIV protease inhibitor, NFV, was loaded into PLGA nanoparticles by double emulsion/solvent evaporation method; and nanoparticles were characterized for physicochemical characteristics including morphology and intracellular uptake. Their anti-cancer efficacy in NSCLC was assessed by in vitro assays including cytotoxicity, cellular migration, colony formation; and 3D spheroid culture mimicking in-vivo tumor microenvironment. Studies were also conducted to elucidate effects on molecular pathways including apoptosis, autophagy, and endoplasmic stress. NFV loaded PLGA nanoparticles (NPs) were found to have particle size: 191.1 ± 10.0nm, zeta potential: -24.3 ± 0.9mV, % drug loading: 2.5 ± 0.0%; and entrapment efficiency (EE): 30.1 ± 0.5%. NFV NP inhibited proliferation of NSCLC cells compared to NFV and exhibited significant IC50 reduction. From the caspase-dependent apoptosis assays and western blot studies (upregulation of ATF3), it was revealed that NFV NP significantly induced ER stress marker ATF3, cleaved PARP and further caused autophagy inhibition (LC3BII upregulation) leading to increased cellular death. In addition, NFV NP were found to be more efficacious in penetrating solid tumors in ex-vivo studies compared to plain NFV. Nelfinavir, a lead HIV protease inhibitor can be repositioned as a NSCLC therapeutic through nanoparticulate delivery. Given its ability to induce apoptosis and efficient tumor penetration capability, NFV loaded PLGA nanoparticulate systems provide a promising delivery system in NSCLC treatment.

The anti-HIV drug nelfinavir mesylate (Viracept) is a potent inhibitor of cell fusion caused by the SARSCoV-2 spike (S) glycoprotein warranting further evaluation as an antiviral against COVID-19 infections.

Severe acute respiratory syndrome coronavirus‐2 (SARS CoV‐2) is the causative agent of the coronavirus disease‐2019 (COVID‐19) pandemic. Coronaviruses enter cells via fusion of the viral envelope with the plasma membrane and/or via fusion of the viral envelope with endosomal membranes after virion endocytosis. The spike (S) glycoprotein is a major determinant of virus infectivity. Herein, we show that the transient expression of the SARS CoV‐2 S glycoprotein in Vero cells caused extensive cell fusion (formation of syncytia) in comparison to limited cell fusion caused by the SARS S glycoprotein. Both S glycoproteins were detected intracellularly and on transfected Vero cell surfaces. These results are in agreement with published pathology observations of extensive syncytia formation in lung tissues of patients with COVID‐19. These results suggest that SARS CoV‐2 is able to spread from cell‐to‐cell much more efficiently than SARS effectively avoiding extracellular neutralizing antibodies. A systematic screening of several drugs including cardiac glycosides and kinase inhibitors and inhibitors of human immunodeficiency virus (HIV) entry revealed that only the FDA‐approved HIV protease inhibitor, nelfinavir mesylate (Viracept) drastically inhibited S‐n‐ and S‐o‐mediated cell fusion with complete inhibition at a 10‐μM concentration. In‐silico docking experiments suggested the possibility that nelfinavir may bind inside the S trimer structure, proximal to the S2 amino terminus directly inhibiting S‐n‐ and S‐o‐mediated membrane fusion. Also, it is possible that nelfinavir may act to inhibit S proteolytic processing within cells. These results warrant further investigations of the potential of nelfinavir mesylate to inhibit virus spread at early times after SARS CoV‐2 symptoms appear.

Antitumorigenic action of nelfinavir: Effects on multiple myeloma and hematologic malignancies (Review).

Protease inhibitors (PIs) inhibit HIV‑1 and HIV‑2 proteases, impeding virus replication and liberation of viral elements from infected cells. In human immunodeficiency virus(HIV) subjects receiving PI‑based treatment, an impressive decrease in the amount of HIV‑associated cancers, unconnected to viral burden or CD4 amount was observed. Research has reported that PIs have influence on cancer proliferation, spread, and survival as an effect on endoplasmic reticulum stress, proteasome, NF‑κB and Akt signalling. Nelfinavir(NFV) is a nonpeptidic PI that functions by connecting to the catalytic site of the HIV protease, thus stopping the cleavage of viral polyprotein into complete, operative proteins that are fundamental for viral survival. NFV, currently not frequently employed for antiretroviral treatment, has demonstrated noteworthy off target effects in tumor patients with or without HIV disease. NFV appears to cause cell death in tumor cells by different mechanisms, which include necrosis, apoptosis and autophagy. In this review, data from preclinical research and clinical trials are reported and the mechanisms of action of NFV and their results in the treatment of hematologic malignancies, such as acute myeloid leukemia, chronic lymphoid leukemia, and diffuse large Bcell lymphoma, and especially in patients with multiple myeloma are examined. In the future, experimental studies may help identify the role of NFV in cancer treatment and may promote the application of this drug into daily clinical practice.

Nanosponge as a drug delivery system

Nanosponges are colloidal carriers that potentially improve the aqueous solubility of poorly water-soluble drug, retarding its release, improving bioavailability, enhancing physical and chemical stability and decrease skin irritation as well. In addition, hydrogels and Nano-sponges are assumed to be excellent applicants for controlled release devices, bio-adhesive devices or target devices of therapeutic agents. Nanosponges may also be used to protect encapsulated molecules from light or from chemical and enzyme induced degradation, e.g. encapsulating 5-fluorouracile, a light-sensitive drug, in Nano-sponges protect it from light degradation, as well shelf life prolongation of camptothecin on encapsulation on Nano-sponges. Nano-sponges also protected the lactone ring from opening due to its high inclusion abilities, thereby increasing stability. Moreover, Nano-sponges can be used to design modified release product to provide slow, continuous delivery of the drug over the entire dosing interval. This makes it possible to decrease the dose administered, change the pharmacokinetic profile, and decrease side effects. Flurbiprofen, Doxorubicin were released slowly when incorporated in β-CD Nano-sponges. As well Nelfinavir mesylate, a protease inhibitor with low bioavailability, used to treat HIV infections, loaded Nano-sponges was prepared to enhance the solubility of the drug with slow release from Nano-sponges than from a β-CD complex.

Synthesis of the compound 4-Coumarinyl 4-methoxybenzoate and molecular docking study against COVID-19

In this study, a coumarin derivative 4-coumarinyl 4-methoxybenzoate was synthesized. Mold / structure. The data of the synthesized compounds were determined to be compatible with IR, 13C and 1H NMR. The COVID-19 activity of the coumarin-derived compound that I synthesized with molecular placement studies was revealed. In addition, these results have been previously studied and compared to Nelfinavir (NFV), which is declared promising for this activity. There is an urgent need for a drug to be used against COVID-19, which negatively affects human life worldwide and causes pandemic leading to the death of many people. The activity potentials of this compound was further evaluated through molecular docking studies with AutoDock4 and AutoDock Vina softwares.

Abstract NT-111: THE ANTIPROGESTIN/ANTIGLUCOCORTICOID MIFEPRISTONE AND THE HIV PROTEASE INHIBITOR NELFINAVIR CAUSE ENDOPLASMIC RETICULUM STRESS AND POTENTIATE THE TOXICITY OF PROTEASOME INHIBITION IN HIGH-GRADE SEROUS EPITHELIAL OVARIAN CANCER CELLS

Abstract Epithelial ovarian cancer (EOC) is a fatal disease due to late diagnosis and lack of effective long-term treatment(s). Since the introduction of debulking surgery and platinum (Pt)-taxane (Tx) therapy over 30 yrs. ago, there has been no significant breakthrough impacting the overall survival of these patients. Though over 70% of diagnosed women respond to front-line standard of care with remission, the disease hides as microscopic (minimal residual) within the abdominal cavity for about 18-24 months (mo.), recurring thereafter with a phenotype usually not responsive to current chemotherapeutic agents. As patients are left without treatment between remission and recurrence, our research initiative is to develop a consolidation therapy for chronic use after standard of care. In this work we explored whether such consolidation therapy could be developed from two simultaneous strategies: proteasome inhibition and aggravation of the stress of the endoplasmic reticulum (ER). The rationale for this combination therapy is that malignant cells are more susceptible to the toxicity of proteasome inhibition and operate with increased expression of ER stress-related proteins—coined as unfolded protein response (UPR) addiction—allowing cancer cells to survive in a hostile environment of reduced nutrients, acidosis, energy deficiency, and low oxygen tension (hypoxia). We hypothesized that simultaneous ER stress aggravation and blockage of the proteolytic capacity of the proteasome should cause sufficient ER stress to tilt cells to a death fate. We report that antiprogestin/antiglucocorticoid mifepristone (MF) as well as HIV protease inhibitor nelfinavir (NFV) enhanced the stress of the ER in EOC cells of high-grade serous origin (HGSOC), which is the most aggressive subtype of EOC, represents the majority of cases of EOC, and causes 2/3 of all deaths from this disease. We demonstrated, in HGSOC cells, that both MF and NFV cause cell cycle arrest associated with increased expression of cyclin dependent kinase inhibitor p27kip1, while triggering the UPR in a dose-dependent manner assessed by increased subrogate ER stress biomarkers GRP78, ATF4, and CHOP. We also discovered that blocking the ubiquitin proteasome system (UPS) using cytostatic concentrations of the proteasome inhibitor bortezomib (BZ) causes accumulation of poly-ubiquitinated proteins and further activation of the UPR. More importantly, when we combined BZ with either MF or NFV, we observed a potentiation of the action of BZ leading to EOC lethality. These results suggest that targeting the ER stress-associated protein quality control machinery with either an antiprogestin/antiglucocorticoid agent or an HIV protease inhibitor may provide an alternative chronic treatment approach against HGSOC after standard of care. In addition, the data provide the rationale for rapid repurposing to treating HGSOC, of three drugs clinically approved for other uses, as their systemic toxicities have already been assessed. Citation Format: Mahbuba Subeha, Lei Zhang, Alicia Goyeneche, Carlos Telleria. THE ANTIPROGESTIN/ANTIGLUCOCORTICOID MIFEPRISTONE AND THE HIV PROTEASE INHIBITOR NELFINAVIR CAUSE ENDOPLASMIC RETICULUM STRESS AND POTENTIATE THE TOXICITY OF PROTEASOME INHIBITION IN HIGH-GRADE SEROUS EPITHELIAL OVARIAN CANCER CELLS [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-111.

Lymphocyte-sparing effect of stereotactic body radiation therapy compared to conventional fractionated radiation therapy in patients with locally advanced pancreatic cancer

BackgroundConventionally fractionated (CF) radiation therapy (RT) has been associated with lymphopenia, leading to compromised overall survival (OS) in cancer patients. It currently remains unknown if stereotactic body (SB) RT induces lymphopenia to the same degree. The aim of this study is to determine if SBRT with either chemotherapy (CMT) (Fluorouracil (5FU) or capecitabine) or Nelfinavir (NFV) to pancreatic adenocarcinoma induces lymphopenia to the same degree as CFRT with 5FU or capecitabine and how any associated difference affects patient survival outcomes.MethodsMedical records of pancreatic adenocarcinoma patients treated with induction CMT followed by RT with concurrent CMT or NFV were reviewed. Patients with total lymphocyte counts (TLCs) available both prior to and following initiation of RT were included. Three groups were identified: CFRT/CMT, SBRT/CMT, and SBRT/NFV. Median delivered RT doses for CFRT and SBRT were 50.4 Gy in 1.8 Gy fractions and 35 Gy in 7 Gy fractions, respectively. TLCs from day 0 (the first day of RT) to 40 were recorded and analyzed using the Kruskal-Wallis test with p-values adjusted with Bonferroni’s method. Linear regressions were utilized to estimate the slope of TLCs as it changes with time and survival analysis was performed via Kaplan-Meier plots.ResultsOne hundred patients were identified (28 CFRT/CMT, 27 SBRT/CMT, 45 SBRT/NFV). Median pre-RT TLCs were not different among groups. Median lowest TLCs were significantly lower (p < 0.0001) and median TLCs reduction over time were significantly greater (p < 0.0001) in the CFRT group than SBRT groups. There was no difference in lowest TLCs or TLCs reduction over time between SBRT groups. Across all groups, the median time to lowest TLCs was similar. Survival analysis revealed no significant difference in median OS between SBRT and CFRT groups. However, in patients with surgery, Median OS for patients with SBRT/CMT was significantly higher than in those with SBRT/NFV (p = 0.03).ConclusionsCompared to CFRT, SBRT is associated with less lymphopenia. Further study of the effect of radiation technique on immune status is warranted.

Nelfinavir interacts with mitochondrial VDACs and disrupts oxidative phosphorylation in proteasome inhibitor resistant myeloma

Proteasome inhibitor (PI) resistance remains a problem in multiple myeloma (MM) therapy. Metabolic reprogramming towards oxidative phosphorylation (OXPHOS) and high mitochondrial ATP supply provide proteasome inhibitor resistance (Tsvetkov, NatureChemBiol 2019). The anti-HIV drug nelfinavir (NFV) overcomes PI-resistance in combination with bortezomib. We identified the molecular targets of NFV and addressed the effect of NFV on the metabolic reprogramming of PI-resistant MM.

Clinical Outcomes of the HIV Protease Inhibitor Nelfinavir With Concurrent Chemoradiotherapy for Unresectable Stage IIIA/IIIB Non–Small Cell Lung Cancer

Local failure after chemoradiotherapy (CT-RT) significantly contributes to mortality in patients with locally advanced non-small cell lung cancer (LA-NSCLC). One approach to improve local control is through targeted radiosensitization of the tumor. To evaluate the dose-limiting toxic effects, maximally tolerated dose, and recommended phase 2 dose of the protease inhibitor nelfinavir mesylate, administered concurrently with CT-RT in patients with LA-NSCLC, and, in the phase 2 portion of the study, to estimate the objective response rate, local and distant failure rates, and overall survival. This prospective, open-label, single-group, single-institution phase 1/2 trial tested the oral protease inhibitor nelfinavir in combination with concurrent CT-RT in 35 patients aged 18 to 89 years with biopsy-confirmed unresectable stage IIIA/IIIB LA-NSCLC and a minimum Karnofsky performance status from June 29, 2007, to February 22, 2012, with an analysis date of May 9, 2017. Median follow-up for all patients was 6.8 years, with a minimum 5 years of follow-up for all survivors. Oral nelfinavir mesylate, 625 mg, twice daily or 1250 mg, twice daily was administered for 7 to 14 days before and during concurrent CT-RT. Graded toxic effects, overall survival, local failure, distant failure, objective response rate, and progression-free survival as measured by Response Evaluation Criteria in Solid Tumors, version 1.1. Thirty-five patients (16 women and 19 men; median age, 60 years [range, 39-79 years]) enrolled and met protocol-specified criteria for adherence, with 5 at a dose of 625 mg twice daily and 30 at a dose of 1250 mg twice daily. No dose-limiting toxic effects were observed. No grade 4 or higher nonhematologic toxic effects were observed. Thirty-three of the 35 patients had evaluable posttreatment computed tomographic scans, with an objective response rate of 94% (31 of 33; 95% CI, 86%-100%). The cumulative incidence of local failure was 39% (95% CI, 30.5%-47.5%). Median progression-free survival was 11.7 months (95% CI, 6.2-17.1 months). Median overall survival for all patients was 41.1 months (95% CI, 19.0-63.1 months); the 5-year mean (SE) overall survival rate was 37.1% (8.2%). This study suggests that nelfinavir administered with concurrent CT-RT is associated with acceptable toxic effects and a promising objective response rate, local failure, progression-free survival, and overall survival in unresectable LA-NSCLC. These data suggest that nelfinavir may enhance the efficacy of standard CT-RT in this disease. Additional testing in the randomized phase 3 setting should be conducted to establish the improvement associated with nelfinavir with concurrent CT-RT. ClinicalTrials.gov identifier: NCT00589056.

The mechanism for differential effect of nelfinavir and indinavir on collagen metabolism in human skin fibroblasts.

The mechanism for differential effects of human immune deficiency virus protease inhibitors (HIVPIs), nelfinavir (NEL) and indinavir (IND) on collagen metabolism disturbances was studied in human skin fibroblasts. It has been considered that HIVPIs-dependent deregulation of collagen biosynthesis involves prolidase (an enzyme providing proline for collagen biosynthesis), glutamine (Gln) (a substrate for proline biosynthesis), nuclear factor-κB (NF-κB) (a transcription factor that inhibit expression of type I collagen genes), β1 integrin receptor and Akt signalling. It was found that NEL impaired collagen biosynthesis and the process was more pronounced in the presence of Gln, while IND stimulated collagen biosynthesis. NEL-dependent inhibition of collagen biosynthesis was accompanied by massive intracellular accumulation of type I collagen, while IND slightly induced this process. This effect of NEL was reversed by ascorbic acid but not N-acetylcysteine. The mechanism for the NEL-dependent defect in collagen metabolism was found at the level of prolidase activity, β1 integrin signalling and NF-κB. NEL inhibited expression of β1 integrin receptor, Akt and ERK1/2 and increased expression of p65 NF-κB. However, inhibitors of p65 NF-κB did not prevent NEL-dependent inhibition of collagen biosynthesis suggesting that this transcription factor is not involved in studied mechanism. Using PI3K inhibitor wortmannin that prevent phosphorylation of Akt revealed that NEL-dependent inhibition of Akt results in inhibition of collagen biosynthesis. The data suggest that differential effect of NEL and IND on collagen metabolism involves NEL-dependent down-regulation of Akt signalling and proline availability for collagen biosynthesis.

Phase I trial of nelfinavir added to cisplatin chemotherapy with concurrent pelvic radiation for locally advanced cervical cancer

Objective: Despite platinum-based chemoradiation (C-XRT), 40%–50% of women with locally advanced cervical cancer will die from their disease. Nelfinavir (NFV), a protease inhibitor, has been shown to target the Akt pathway sensitizing cancer cells to chemoradiation. The objective of this phase I trial was to evaluate the safety and tolerability and identify the recommended phase II dose (RP2D) of NFV in combination with standard concurrent C-XRT in locally advanced cervical cancer.

Solid-phase extraction combined with dispersive liquid-liquid microextraction/HPLC-UV as a sensitive and efficient method for extraction, pre-concentration and simultaneous determination of antiretroviral drugs nevirapine, efavirenz and nelfinavir in pharmaceutical formulations and biological samples

Therapeutic drug monitoring (TDM) of antiretroviral drugs requires accurate and precise analysis of their trace amounts in plasma samples. Solid-phase extraction (SPE) coupled with dispersive liquid-liquid microextraction based on solidification of floating organic drop (DLLME-SFO) was introduced as a simple and sensitive method for extraction, pre-concentration and simultaneous determination of efavirenz (EFV), nelfinavir (NFV) and nevirapine (NVP) in human plasma and pharmaceutical formulations by using high performance liquid chromatography (HPLC) with UV detection. A response surface methodology (RSM) based on central composite design (CCD) was used for optimizing the main variables in the extraction procedure. Under the optimum experimental conditions, the calibration curves were linear in the range of 0.1–400 ng mL−1 with correlation coefficients from 0.9982 to 0.9997 and limits of detection (at S/N = 3) of 0.03 to 0.07 ng mL−1. Moreover the intra-day and inter-day precision (RSD%) were in the range of 2.2–4.2% and 3.1–5.2%, respectively. The enrichment factors and relative recoveries of the anti-HIV drugs were 459–1507 and 93.7–105.4%. The method was successfully applied to the simultaneous determination of the trace amounts of the antiretroviral drugs in blood plasma and pharmaceutical formulations.