Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active andr... more Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active androgen dihydrotestosterone (DHT) which is involved in many androgen dependent disorders, such as androgenic alopecia, benign prostatic hyperplasia and acne. The method for screening for S5αR inhibition is key in finding new antagonists. In this study, the label-free S5αR inhibitory assay using LC-MS was developed. S5αR type 1 enzyme was obtained from LNCaP prostate cancer cells. The enzymatic assay was optimized for enzyme-substrate (testosterone) concentration, NADPH-cofactor concentration, solvent tolerance, enzyme activity stability and incubation time. The developed assay was validated by measuring the signal to background ratio (S/B), the signal to noise ratio (S/N), the signal window (SW) and the zeta factor Z′ in accordance with published bioassay guidelines. The enzymatic reaction was performed in 96well plates and DHT formation was determined by LC-MS. S/B, S/N, SW and Z′ factor were well above acceptable criteria and the reproducibility was good using Z′ factor other 3 days and further validated by dutasteride and finasteride inhibition. The method was successfully applied to quantify S5αR inhibitory activity of some Thai herbal extracts. Two plant extracts, Impatiens balsaminaL. and Curcuma longaL. showed IC 50 at 5.4±0.2 and 9.0±1.2 µgmL-1 and are therefore promising sources of new S5αR inhibitors. The assay has high selectability and reproducibility and suited to medium throughput screening required by phytochemistry.
Journal of the American Pharmacists Association, 2020
Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes o... more Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes on pain and adverse euphoria events. Data sources: A systematic search was performed in PubMed, ScienceDirect, ClincalTrials.gov, Scopus, Cochrane Library, and Embase from inception until June 2017. Study selection: Randomized controlled trials investigating the effects of cannabis or cannabinoids on pain reduction. Data extraction: Two reviewers extracted and assessed the quality of studies by means of Cochrane risk of bias. Standardized mean difference (SMD) was calculated. Random-effects model was undertaken to pool the treatment effects. Results: A total of 25 studies involving 2270 patients were included. We found that delta-9tetrahydrocannabinol/cannabidiol (THC/CBD) (oromucosal route), THC (oromucosal route), and standardized dried cannabis (with THC; SCT; inhalation route) could reduce neuropathic pain score (SMD À0.41, 95% CI À0.7 to À0.1; À0.61, 95% CI À1.2 to À0.02; and À0.77, 95% CI À1.4 to À0.2; respectively). For nociceptive pain, only standardized cannabis extract (with THC; SCET) via oral route could reduce pain score (SMD À1.8, 95% C; À2.4 to À1.2). In cancer pain, THC/CBD via oromucosal route and THC via oral or oromucosal route could reduce pain score (SMD À0.7, 95% CI À1.2 to À0.2; and À2.1, 95% CI À2.8 to À1.4; respectively). No study was observed for THC/CBD via oral route or inhalation or THC via inhalation for cancer and nociceptive pain, SCET via oromucosal route or inhalation for neuropathic and cancer pain, THC via oromucosal route for nociceptive pain, and SCT via oromucosal or oral route for neuropathic, cancer, and nociceptive pain. Statistically significant increased risks of euphoria were observed in THC/CBD (oromucosal), THC (oromucosal), and SCT (inhalation). Conclusion: The use of cannabis and cannabinoids via certain administration routes could reduce different types of pain. Product developers could consider our findings as part of their product design so that the effective route of cannabis and cannabinoids for pain control can be achieved.
Marine sponges have been the source of various metabolites with potent biological activities. In ... more Marine sponges have been the source of various metabolites with potent biological activities. In this study fifteen methanolic extracts of marine sponges, collected off the coast of Tabuhan Island, Banyuwangi, East Java, Indonesia were evaluated in relation to their cholinesterase and 5α-reductase inhibitory activities. The results revealed that the extract of Petrosia sp. inhibited the 5α-reductase enzyme at 100 µg/mL, with 61.21% inhibition, which is slightly lower than the positive control, finasteride, of 76.70%. The results of the cholinesterase inhibitory screening showed that three marine sponges namely, Callyspongia sp., Niphates olemda, and Agelas nakamurai presented notable cholinesterase inhibitory activities. The highest potency was found in A. nakamurai, with an IC50 value of 1.05 µg/mL. All three samples inhibited both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE), however, the extract of N. olemda showed a higher inhibition against AChE compared to BuC...
Journal of the American Pharmacists Association, 2019
Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes o... more Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes on pain and adverse euphoria events. Data sources: A systematic search was performed in PubMed, ScienceDirect, ClincalTrials.gov, Scopus, Cochrane Library, and Embase from inception until June 2017. Study selection: Randomized controlled trials investigating the effects of cannabis or cannabinoids on pain reduction. Data extraction: Two reviewers extracted and assessed the quality of studies by means of Cochrane risk of bias. Standardized mean difference (SMD) was calculated. Random-effects model was undertaken to pool the treatment effects. Results: A total of 25 studies involving 2270 patients were included. We found that delta-9tetrahydrocannabinol/cannabidiol (THC/CBD) (oromucosal route), THC (oromucosal route), and standardized dried cannabis (with THC; SCT; inhalation route) could reduce neuropathic pain score (SMD À0.41, 95% CI À0.7 to À0.1; À0.61, 95% CI À1.2 to À0.02; and À0.77, 95% CI À1.4 to À0.2; respectively). For nociceptive pain, only standardized cannabis extract (with THC; SCET) via oral route could reduce pain score (SMD À1.8, 95% C; À2.4 to À1.2). In cancer pain, THC/CBD via oromucosal route and THC via oral or oromucosal route could reduce pain score (SMD À0.7, 95% CI À1.2 to À0.2; and À2.1, 95% CI À2.8 to À1.4; respectively). No study was observed for THC/CBD via oral route or inhalation or THC via inhalation for cancer and nociceptive pain, SCET via oromucosal route or inhalation for neuropathic and cancer pain, THC via oromucosal route for nociceptive pain, and SCT via oromucosal or oral route for neuropathic, cancer, and nociceptive pain. Statistically significant increased risks of euphoria were observed in THC/CBD (oromucosal), THC (oromucosal), and SCT (inhalation). Conclusion: The use of cannabis and cannabinoids via certain administration routes could reduce different types of pain. Product developers could consider our findings as part of their product design so that the effective route of cannabis and cannabinoids for pain control can be achieved.
Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active andr... more Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active androgen dihydrotestosterone (DHT) which is involved in many androgen dependent disorders, such as androgenic alopecia, benign prostatic hyperplasia and acne. The method for screening for S5αR inhibition is key in finding new antagonists. In this study, the label-free S5αR inhibitory assay using LC-MS was developed. S5αR type 1 enzyme was obtained from LNCaP prostate cancer cells. The enzymatic assay was optimized for enzyme-substrate (testosterone) concentration, NADPH-cofactor concentration, solvent tolerance, enzyme activity stability and incubation time. The developed assay was validated by measuring the signal to background ratio (S/B), the signal to noise ratio (S/N), the signal window (SW) and the zeta factor Z′ in accordance with published bioassay guidelines. The enzymatic reaction was performed in 96well plates and DHT formation was determined by LC-MS. S/B, S/N, SW and Z′ factor were well above acceptable criteria and the reproducibility was good using Z′ factor other 3 days and further validated by dutasteride and finasteride inhibition. The method was successfully applied to quantify S5αR inhibitory activity of some Thai herbal extracts. Two plant extracts, Impatiens balsaminaL. and Curcuma longaL. showed IC 50 at 5.4±0.2 and 9.0±1.2 µgmL-1 and are therefore promising sources of new S5αR inhibitors. The assay has high selectability and reproducibility and suited to medium throughput screening required by phytochemistry.
Anti-androgen can be used in the treatment of benign prostatic hyperplasia, acne, hirsutism, and ... more Anti-androgen can be used in the treatment of benign prostatic hyperplasia, acne, hirsutism, and androgenic alopecia. For the search of anti-androgenic activity through steroid 5-alpha reductase (S5αR) inhibition mechanism, 12 natural analogs from plant origins, i.e., curcumin (1) demethoxycurcumin (2), and bisdemethoxycurcumin (3) isolated from Curcuma longa Linn., compounds 18, 20, 21, 22, 24, and 25 isolated from Curcuma comosa Roxb., amide analogs 29-31 obtained from Bougainvillea spectabilis Willd. together with 21 synthesized analogs were evaluated for S5αR inhibitory activity using liquid chromatography-mass spectrometry assay. The results showed that compounds 1, 2, 4, 5, 6, 7, and 9 possessed S5αR inhibitory activity and compounds 1, 4, and 5 were the most potent (IC 50 of 13.4 ± 0.4, 15.3 ± 3.1 and 8.9 ± 0.9 µM, respectively). This suggests that the unsaturated enone moiety in the chain linked between two aromatic rings of curcumin analog was imperative to the activity. Moreover, the m-methoxyl and p-hydroxyl substitutions in aromatic region of 1,6-heptadiene-3,5-dione linker were necessary. The cytotoxic effect on androgen-dependent cell, human dermal papilla was investigated to obtain safety information profile. We found that 1,6-heptadiene-3,5-dione linker was important for safety. This work stated that antiandrogen activity of curcumin analogs was through S5αR inhibition mechanism and the information might lead to further design of new curcumin analogs with improved potency and safety.
Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active andr... more Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active androgen dihydrotestosterone (DHT) which is involved in many androgen dependent disorders, such as androgenic alopecia, benign prostatic hyperplasia and acne. The method for screening for S5αR inhibition is key in finding new antagonists. In this study, the label-free S5αR inhibitory assay using LC-MS was developed. S5αR type 1 enzyme was obtained from LNCaP prostate cancer cells. The enzymatic assay was optimized for enzyme-substrate (testosterone) concentration, NADPH-cofactor concentration, solvent tolerance, enzyme activity stability and incubation time. The developed assay was validated by measuring the signal to background ratio (S/B), the signal to noise ratio (S/N), the signal window (SW) and the zeta factor Z′ in accordance with published bioassay guidelines. The enzymatic reaction was performed in 96well plates and DHT formation was determined by LC-MS. S/B, S/N, SW and Z′ factor were well above acceptable criteria and the reproducibility was good using Z′ factor other 3 days and further validated by dutasteride and finasteride inhibition. The method was successfully applied to quantify S5αR inhibitory activity of some Thai herbal extracts. Two plant extracts, Impatiens balsaminaL. and Curcuma longaL. showed IC 50 at 5.4±0.2 and 9.0±1.2 µgmL-1 and are therefore promising sources of new S5αR inhibitors. The assay has high selectability and reproducibility and suited to medium throughput screening required by phytochemistry.
Journal of the American Pharmacists Association, 2020
Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes o... more Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes on pain and adverse euphoria events. Data sources: A systematic search was performed in PubMed, ScienceDirect, ClincalTrials.gov, Scopus, Cochrane Library, and Embase from inception until June 2017. Study selection: Randomized controlled trials investigating the effects of cannabis or cannabinoids on pain reduction. Data extraction: Two reviewers extracted and assessed the quality of studies by means of Cochrane risk of bias. Standardized mean difference (SMD) was calculated. Random-effects model was undertaken to pool the treatment effects. Results: A total of 25 studies involving 2270 patients were included. We found that delta-9tetrahydrocannabinol/cannabidiol (THC/CBD) (oromucosal route), THC (oromucosal route), and standardized dried cannabis (with THC; SCT; inhalation route) could reduce neuropathic pain score (SMD À0.41, 95% CI À0.7 to À0.1; À0.61, 95% CI À1.2 to À0.02; and À0.77, 95% CI À1.4 to À0.2; respectively). For nociceptive pain, only standardized cannabis extract (with THC; SCET) via oral route could reduce pain score (SMD À1.8, 95% C; À2.4 to À1.2). In cancer pain, THC/CBD via oromucosal route and THC via oral or oromucosal route could reduce pain score (SMD À0.7, 95% CI À1.2 to À0.2; and À2.1, 95% CI À2.8 to À1.4; respectively). No study was observed for THC/CBD via oral route or inhalation or THC via inhalation for cancer and nociceptive pain, SCET via oromucosal route or inhalation for neuropathic and cancer pain, THC via oromucosal route for nociceptive pain, and SCT via oromucosal or oral route for neuropathic, cancer, and nociceptive pain. Statistically significant increased risks of euphoria were observed in THC/CBD (oromucosal), THC (oromucosal), and SCT (inhalation). Conclusion: The use of cannabis and cannabinoids via certain administration routes could reduce different types of pain. Product developers could consider our findings as part of their product design so that the effective route of cannabis and cannabinoids for pain control can be achieved.
Marine sponges have been the source of various metabolites with potent biological activities. In ... more Marine sponges have been the source of various metabolites with potent biological activities. In this study fifteen methanolic extracts of marine sponges, collected off the coast of Tabuhan Island, Banyuwangi, East Java, Indonesia were evaluated in relation to their cholinesterase and 5α-reductase inhibitory activities. The results revealed that the extract of Petrosia sp. inhibited the 5α-reductase enzyme at 100 µg/mL, with 61.21% inhibition, which is slightly lower than the positive control, finasteride, of 76.70%. The results of the cholinesterase inhibitory screening showed that three marine sponges namely, Callyspongia sp., Niphates olemda, and Agelas nakamurai presented notable cholinesterase inhibitory activities. The highest potency was found in A. nakamurai, with an IC50 value of 1.05 µg/mL. All three samples inhibited both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE), however, the extract of N. olemda showed a higher inhibition against AChE compared to BuC...
Journal of the American Pharmacists Association, 2019
Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes o... more Objectives: To determine the effects of cannabis, cannabinoids, and their administration routes on pain and adverse euphoria events. Data sources: A systematic search was performed in PubMed, ScienceDirect, ClincalTrials.gov, Scopus, Cochrane Library, and Embase from inception until June 2017. Study selection: Randomized controlled trials investigating the effects of cannabis or cannabinoids on pain reduction. Data extraction: Two reviewers extracted and assessed the quality of studies by means of Cochrane risk of bias. Standardized mean difference (SMD) was calculated. Random-effects model was undertaken to pool the treatment effects. Results: A total of 25 studies involving 2270 patients were included. We found that delta-9tetrahydrocannabinol/cannabidiol (THC/CBD) (oromucosal route), THC (oromucosal route), and standardized dried cannabis (with THC; SCT; inhalation route) could reduce neuropathic pain score (SMD À0.41, 95% CI À0.7 to À0.1; À0.61, 95% CI À1.2 to À0.02; and À0.77, 95% CI À1.4 to À0.2; respectively). For nociceptive pain, only standardized cannabis extract (with THC; SCET) via oral route could reduce pain score (SMD À1.8, 95% C; À2.4 to À1.2). In cancer pain, THC/CBD via oromucosal route and THC via oral or oromucosal route could reduce pain score (SMD À0.7, 95% CI À1.2 to À0.2; and À2.1, 95% CI À2.8 to À1.4; respectively). No study was observed for THC/CBD via oral route or inhalation or THC via inhalation for cancer and nociceptive pain, SCET via oromucosal route or inhalation for neuropathic and cancer pain, THC via oromucosal route for nociceptive pain, and SCT via oromucosal or oral route for neuropathic, cancer, and nociceptive pain. Statistically significant increased risks of euphoria were observed in THC/CBD (oromucosal), THC (oromucosal), and SCT (inhalation). Conclusion: The use of cannabis and cannabinoids via certain administration routes could reduce different types of pain. Product developers could consider our findings as part of their product design so that the effective route of cannabis and cannabinoids for pain control can be achieved.
Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active andr... more Steroid 5α-reductase (S5αR) plays an important role in metabolizing testosterone into active androgen dihydrotestosterone (DHT) which is involved in many androgen dependent disorders, such as androgenic alopecia, benign prostatic hyperplasia and acne. The method for screening for S5αR inhibition is key in finding new antagonists. In this study, the label-free S5αR inhibitory assay using LC-MS was developed. S5αR type 1 enzyme was obtained from LNCaP prostate cancer cells. The enzymatic assay was optimized for enzyme-substrate (testosterone) concentration, NADPH-cofactor concentration, solvent tolerance, enzyme activity stability and incubation time. The developed assay was validated by measuring the signal to background ratio (S/B), the signal to noise ratio (S/N), the signal window (SW) and the zeta factor Z′ in accordance with published bioassay guidelines. The enzymatic reaction was performed in 96well plates and DHT formation was determined by LC-MS. S/B, S/N, SW and Z′ factor were well above acceptable criteria and the reproducibility was good using Z′ factor other 3 days and further validated by dutasteride and finasteride inhibition. The method was successfully applied to quantify S5αR inhibitory activity of some Thai herbal extracts. Two plant extracts, Impatiens balsaminaL. and Curcuma longaL. showed IC 50 at 5.4±0.2 and 9.0±1.2 µgmL-1 and are therefore promising sources of new S5αR inhibitors. The assay has high selectability and reproducibility and suited to medium throughput screening required by phytochemistry.
Anti-androgen can be used in the treatment of benign prostatic hyperplasia, acne, hirsutism, and ... more Anti-androgen can be used in the treatment of benign prostatic hyperplasia, acne, hirsutism, and androgenic alopecia. For the search of anti-androgenic activity through steroid 5-alpha reductase (S5αR) inhibition mechanism, 12 natural analogs from plant origins, i.e., curcumin (1) demethoxycurcumin (2), and bisdemethoxycurcumin (3) isolated from Curcuma longa Linn., compounds 18, 20, 21, 22, 24, and 25 isolated from Curcuma comosa Roxb., amide analogs 29-31 obtained from Bougainvillea spectabilis Willd. together with 21 synthesized analogs were evaluated for S5αR inhibitory activity using liquid chromatography-mass spectrometry assay. The results showed that compounds 1, 2, 4, 5, 6, 7, and 9 possessed S5αR inhibitory activity and compounds 1, 4, and 5 were the most potent (IC 50 of 13.4 ± 0.4, 15.3 ± 3.1 and 8.9 ± 0.9 µM, respectively). This suggests that the unsaturated enone moiety in the chain linked between two aromatic rings of curcumin analog was imperative to the activity. Moreover, the m-methoxyl and p-hydroxyl substitutions in aromatic region of 1,6-heptadiene-3,5-dione linker were necessary. The cytotoxic effect on androgen-dependent cell, human dermal papilla was investigated to obtain safety information profile. We found that 1,6-heptadiene-3,5-dione linker was important for safety. This work stated that antiandrogen activity of curcumin analogs was through S5αR inhibition mechanism and the information might lead to further design of new curcumin analogs with improved potency and safety.
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