Saquinavir

合成路线和反应条件： 沙奎那韦的合成涉及多个步骤，从关键中间体的制备开始。一种方法包括使用甲磺酰氯在三乙胺和甲苯存在下，在氮气气氛中对 (1S, 2S)-(1-苄基-3-氯-2-羟丙基) 叔丁基氨基甲酸酯进行甲磺酰化。然后，将该中间体在 18-冠醚-6 和甲苯存在下与金属乙酸酯反应，得到 (1S, 2R)-(1-苄基-3-氯-2-乙酸丙基) 叔丁基氨基甲酸酯。 与 KOH、THF 和乙醇进一步反应得到 (2R, 3S)-1,2-环氧-3-叔丁氧羰基氨基-4-苯基丁烷 .

工业生产方法： 沙奎那韦的工业生产通常涉及使用优化反应条件的大规模合成，以确保高收率和纯度。 该工艺包括使用差示扫描量热法 (DSC)、粉末 X 射线衍射 (PXRD) 和傅里叶变换红外光谱 (FT-IR) 等先进技术进行表征和质量控制 .

反应类型： 沙奎那韦会经历各种化学反应，包括氧化、还原和取代反应。这些反应对于其代谢和从体内消除至关重要。

常见试剂和条件：

氧化： 使用过氧化氢或高锰酸钾等试剂可以氧化沙奎那韦。

还原： 还原反应可能涉及硼氢化钠或氢化铝锂等试剂。

取代： 取代反应通常使用亚硫酰氯或三溴化磷等卤化试剂。

主要产物： 从这些反应形成的主要产物包括各种代谢物，这些代谢物主要通过粪便和尿液排泄 .

HIV Treatment

Saquinavir has revolutionized the treatment of HIV/AIDS since its approval in 1995. It is primarily used in combination with other antiretroviral agents as part of Highly Active Antiretroviral Therapy (HAART). Its mechanism involves inhibiting the HIV protease enzyme, which is crucial for viral replication.

Clinical Efficacy:

• This compound has shown significant viral load reduction in patients when used alongside nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) .

Cancer Treatment

Recent studies have indicated that this compound may serve as an effective treatment for various cancers, particularly anal cancer and cervical cancer.

Anal Cancer

A study highlighted this compound's ability to reduce tumor spheroid growth in a transgenic mouse model of anal cancer associated with HPV. The results showed a statistically significant reduction in tumor growth compared to control groups, suggesting its potential as an adjuvant therapy alongside existing protocols .

Cervical Cancer

This compound has also been investigated for its effects on cervical cancer cells. It demonstrated inhibitory effects on cell proliferation and invasion, indicating potential pathways for therapeutic intervention .

COVID-19 Treatment

This compound's antiviral properties have prompted research into its effectiveness against SARS-CoV-2. Molecular docking studies revealed that this compound can bind to the 3CLpro enzyme of the virus, inhibiting its activity. This suggests a possible role in treating COVID-19, especially given its low IC50 value of approximately 9.92 μM .

Tuberculosis Therapy

Emerging evidence supports this compound's use as a host-directed therapy for tuberculosis (TB). Research indicates that this compound enhances the intracellular killing of Mycobacterium tuberculosis (Mtb) in infected macrophages while improving antigen presentation and T-cell responses . This dual action positions it as a promising adjunct treatment for TB.

Microbicide Development

This compound has been explored as a candidate microbicide to prevent HIV transmission. Studies demonstrated its ability to inhibit early events associated with HIV infection in various cell types, including CD4 T cells and macrophages. This positions this compound as a potential preventive measure against HIV in at-risk populations .

Summary of Applications

沙奎那韦通过抑制 HIV-1 蛋白酶发挥其抗病毒活性。该酶对于将多蛋白裂解成功能性病毒蛋白至关重要。 通过阻断这种酶，沙奎那韦阻止了病毒的成熟，导致形成非感染性病毒颗粒 . 主要分子靶标是 HIV-1 蛋白酶，途径涉及病毒复制和组装的破坏 .

类似化合物：

利托那韦： 另一种蛋白酶抑制剂，与其他抗逆转录病毒药物联用。它以其抑制其他蛋白酶抑制剂的代谢，从而增强其有效性而闻名。

茚地那韦： 另一种靶向 HIV-1 蛋白酶的蛋白酶抑制剂，但其药代动力学特性不同。

洛匹那韦： 通常与利托那韦联用，它具有类似的作用机制，但耐药谱不同。

沙奎那韦的独特性： 沙奎那韦是第一个获得批准的蛋白酶抑制剂，与其他抗逆转录病毒疗法相比，其不良反应谱相对较好 . 其独特的结构和与 HIV-1 蛋白酶的结合亲和力使其成为 HIV/AIDS 联合疗法的宝贵组成部分 .

Ritonavir: Another protease inhibitor used in combination with other antiretrovirals. It is known for its ability to boost the effectiveness of other protease inhibitors by inhibiting their metabolism.

Indinavir: A protease inhibitor that also targets the HIV-1 protease but has different pharmacokinetic properties.

Lopinavir: Often used in combination with Ritonavir, it has a similar mechanism of action but different resistance profiles.

Uniqueness of Saquinavir: this compound was the first protease inhibitor to be approved and has a relatively benign adverse effect profile compared to other antiretroviral therapies . Its unique structure and binding affinity to the HIV-1 protease make it a valuable component of combination therapies for HIV/AIDS .

Saquinavir is a protease inhibitor that has been pivotal in the treatment of HIV/AIDS since its introduction. Its biological activity extends beyond antiviral effects, showing potential applications in cancer therapy and other diseases. This article provides a comprehensive overview of the biological activity of this compound, highlighting its mechanisms, efficacy, and case studies.

This compound functions primarily by inhibiting the HIV-1 protease enzyme, which is crucial for the maturation of viral proteins. By preventing this cleavage, this compound effectively reduces viral replication. The compound's limited bioavailability (approximately 4%) necessitates co-administration with ritonavir to enhance its serum concentration and therapeutic efficacy .

Antiviral Activity

• Inhibition of HIV Replication : this compound demonstrates dose-dependent inhibition of HIV-1 replication across various cell types, including CD4 T cells and macrophages . It significantly reduces viral load and improves immune response markers such as HLA class II expression and IFN-γ secretion in infected macrophages .
• Resistance Patterns : Clinical studies indicate that patients with prior exposure to this compound may develop resistance, complicating subsequent treatment with other protease inhibitors like nelfinavir .

Anticancer Potential

Recent studies have explored this compound's role in cancer treatment, particularly in cervical cancer. This compound has been shown to inhibit cell proliferation, invasion, and clonogenicity in HeLa cells at concentrations lower than those required to affect proteasomal activity . This suggests that this compound may exert its effects through alternative oncogenic pathways.

Lipid Metabolism Effects

This compound also impacts lipid metabolism significantly. Research indicates that it inhibits lipoprotein lipase (LPL) activity and lipid synthesis in adipocytes, which may contribute to metabolic complications observed in patients undergoing protease inhibitor therapy . Notably, after removal of this compound, LPL activity partially recovers, indicating a reversible effect on lipid metabolism.

Table 1: Summary of Key Research Findings on this compound

Clinical Implications

A randomized study evaluated the safety and efficacy of a regimen including this compound, ritonavir, and atazanavir over 48 weeks. The results indicated a modest increase in CD4 lymphocyte counts but highlighted adverse effects such as scleral icterus in some patients . These findings underscore the need for careful monitoring during treatment.

Q. Basic Research: What are the primary metabolic pathways of saquinavir, and how do they influence experimental design in pharmacokinetic studies?

This compound is predominantly metabolized by hepatic CYP3A4, with >90% biotransformation occurring via this enzyme . Its first-pass metabolism results in low oral bioavailability (~4%), which is significantly enhanced by co-administration with ritonavir (a CYP3A4 inhibitor) or high-fat meals .
Methodological Guidance :

• Use in vitro liver microsome assays to quantify CYP3A4-mediated metabolism.
• Conduct pharmacokinetic studies in animal/human models with controlled diets (e.g., high-fat vs. fasting) to assess food effects .
• Include CYP3A4 inhibitors/inducers (e.g., ketoconazole, rifampicin) to validate metabolic pathways.

Q. Basic Research: How does this compound interact with P-glycoprotein (P-gp), and what are the implications for cellular uptake studies?

This compound is a substrate of P-gp, an efflux transporter localized on apical surfaces of hepatocytes, renal proximal tubules, and intestinal epithelia . This interaction limits its intracellular accumulation and contributes to variable tissue penetration.
Methodological Guidance :

• Use polarized cell lines (e.g., Caco-2 monolayers) to measure bidirectional transport and P-gp inhibition/induction.
• Quantify intracellular this compound levels via LC-MS/MS in the presence/absence of P-gp inhibitors (e.g., verapamil) .

Q. Advanced Research: How can researchers resolve contradictory data on this compound’s off-target inhibition of cathepsin L versus its lack of effect on proteasome activity in certain models?

This compound inhibits cathepsin L (IC50: 13.4 µM) but shows negligible proteasome inhibition at clinically relevant concentrations (<30 µM) . Discrepancies arise from dose-dependent effects and assay conditions (e.g., exposure time, cell type).
Methodological Guidance :

• Standardize assays using recombinant enzymes (e.g., cathepsin L) and fluorescent substrates to measure inhibition kinetics .
• For proteasome activity, employ cell-based assays with prolonged exposure (≥24 hrs) and higher doses (≥60 µM) to detect modulation .
• Apply statistical tools like two-way ANOVA with Bonferroni correction to compare dose-response curves across cell lines .

Q. Advanced Research: What experimental strategies are recommended to investigate this compound’s potential repurposing for SARS-CoV-2 or cancer therapy?

This compound inhibits SARS-CoV-2 spike protein-mediated cell fusion via cathepsin L blockade and reduces cervical cancer cell proliferation/invasion independently of proteasome modulation .
Methodological Guidance :

• Use pseudovirus entry assays with TMPRSS2/ACE2-expressing cells to quantify SARS-CoV-2 inhibition .
• For cancer studies, perform clonogenic assays and 3D invasion models (e.g., Matrigel) with primary cervical cancer cell lines .
• Validate findings in vivo using xenograft models, monitoring this compound tissue penetration via HPLC .

Q. Advanced Research: How can structural biology elucidate this compound’s resistance profile in HIV protease mutants?

This compound loses efficacy against HIV protease mutants (e.g., G48V) due to disrupted hydrogen bonding, whereas darunavir retains activity . X-ray crystallography reveals differential binding interactions.
Methodological Guidance :

• Solve co-crystal structures of this compound with mutant proteases (e.g., G48V, PRI50V) to map residue-specific interactions .
• Combine molecular dynamics simulations with enzymatic inhibition assays (e.g., fluorescence-based protease activity) to correlate structural changes with resistance .

Q. Advanced Research: What factorial design approaches optimize this compound delivery systems (e.g., transdermal nanoemulsions)?

Self-nanoemulsifying drug delivery systems (SNEDDS) improve this compound bioavailability by enhancing solubility and bypassing first-pass metabolism .
Methodological Guidance :

• Apply Box-Behnken or central composite designs to optimize SNEDDS variables (e.g., oil phase ratio, surfactant concentration) .
• Characterize ex vivo skin permeation using Franz diffusion cells and confocal microscopy for tissue distribution analysis .

Q. Basic Research: How do drug-drug interactions (DDIs) between this compound and CYP3A4 modulators affect experimental outcomes?

This compound’s AUC increases 177% when co-administered with clarithromycin (a CYP3A4 inhibitor), while CYP3A4 inducers (e.g., rifampicin) reduce exposure .
Methodological Guidance :

• Conduct DDI studies in human hepatocyte co-cultures or using physiologically based pharmacokinetic (PBPK) modeling .
• Monitor QT prolongation risks in preclinical models via patch-clamp assays or ECG in animal studies .