• Archives of Pharmacal Research
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• 제28권9호
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• pp.1086-1091
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• 2005

An analytical method was developed for evaluating the cannabidiol (CBO), cannabinol (CBN), ${\Delta}^9-tetrahydrocannabinol$ $({\Delta}^9-THC)$ level in human hair using gas chromatography-mass spectrometry (GC-MS). Hair samples (50mg) were washed with isopropyl alcohol and cut into small fragments (< 1mm). After adding a deuterated internal standard, the hair samples were incubated in 1.0M NaOH for 10 min at $95^{\circ}C$. The analytes from the resulting hydrolyzed samples were extracted using a mixture of n-hexane-ethyl acetate (75:25, v/v). The extracts were then evaporated, derivatized, and injected into the GC-MS. The recovery ranges of CBD, CBN, and ${\Delta}^9-THC$ at three concentration levels were $37.9-94.5\%$ with good correlation coefficients $(r^2>0.9989)$. The intra-day precision and accuracy ranged from $-9.4\%\;to\;17.7\%$, and the inter-day precision and accuracy ranged from $-15.5\%\;to\;14.5\%$, respectively. The limits of detection (LOD) for CBD, CBN, and ${\Delta}^9-THC$ were 0.005, 0.002, and 0.006 ng/mg, respectively. The applicability of this method of analyzing the hair samples from cannabis abusers was demonstrated.

• 약학회지
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• 제17권1호
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• pp.21-26
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• 1973

The constituents of Korean cannabis were studied comparatively with foreign orgins. Especially, tetrahydrocannabinol(THC), the active constituents of cannabis were analyzed by the technique of thin layer chromatography and colorimetry. The results are as follows ; (1) THC content in Korean cannabis is comparatively higher than that in foreign samples. (2) THC is contained most abundantly in male flowers, abundantly in female tops and leaves and some in barks. (3) There is a tendency that the THC content increase gradually with growth of plants, being highest during unripe nad decrease with maturity of the tops. (4) Korean cannabis contains THC, cannabinol, cannabidiol, cannabidiolic acid and other cannandiolic compounds. Distribution of chemical components of Korean cannabis, compared with those of foreign ones, is remarkably different. (5) The THC content of Spanish cannabis cultivated tentatively in Korea is similar to that of Korean cannabis.

• Archives of Pharmacal Research
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• 제28권12호
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• pp.1365-1375
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• 2005

The antiestrogenic effects of marijuana smoke condensate (MSC) and three major cannabinoids, i.e., $\bigtriangleup^{9}$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN), were evaluated using in vitro bioassays, viz., the human breast cancer cell proliferation assay, the recombinant human estrogen receptor (ER) competitive binding assay, and the reporter gene assay. The inhibitory effects on estrogen were also examined using the ethoxyresorufin-O­deethylase (EROD) assay, the aromatase assay, and the 17$\beta$-estradiol ($E_{2}$) metabolism assay. The results showed that MSC induced the antiestrogenic effect via the ER-mediated pathway, while THC, CBD, and CBN did not have any antiestrogenic activity. This suggests that the combined effects of the marijuana smoke components are responsible for the antiestrogenicity of marijuana use. In addition, MSC induced the CYP1A activity and the $E_{2}$ metabolism, but inhibited the aromatase activity, suggesting that the antiestrogenic activity of MSC is also related to the indirect ER-dependent pathway, as a result of the depletion of the in situ $E_{2}$ level available to bind to the ER. In conclusion, pyrogenic products including polycyclic aromatic hydrocarbons (PAHs) in the non-polar fraction, which is the most biologically active fraction among the seven fractions of MSC, might be responsible for the antiestrogenic effect.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.194-199
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• 2002

Many oxidative metabolites of tetrahydrocannabinols (THCs), active components of marijuana, were pharmacologically active, and 11-hydroxy-THCs, 11-oxo-${\Delta}^8$-THC, 7-oxo-${\Delta}^8$-THC, 8$\beta$, 9$\beta$-epoxyhexahydrocannabinol (EHHC), 9$\alpha$, l0$\alpha$-EHHC and 3'-hydroxy-${\Delta}^9$-THC were more active than THC in pharmacological effects such as catalepsy, hypothermia and barbiturate synergism in mice. Cannabidiol (CBD), another major component, was biotransfomred to two novel metabolites, 6-hydroxymethyl-${\Delta}^9$-THC and 3-pentyl-6, 7, 7a, 8, 9, lla-hexahydro-I, 7-dihydroxy-7, 1O-dimethyldibenzo[b, d]oxepin (PHDO) through 8R, 9-epoxy-CBD and 85, 9-epoxy-CBD, respectively. Both metabolites exhibited some pharmacological effects comparable to d9 - THe. Cannabinol (CBN), the other major component, was mainly metabolized to ll-hydroxy-CBN by hepatic microsomes of animals including humans. The pharmacological effects of the metabolite were higher than those of CBN demonstrating that II-hydroxylation of CBN is metabolic activation pathway of the cannabinoid as is the case in THCs. Tolerance and reciprocal cross-tolerance developed to pharmacological effects d8 - THC and ll-hydroxy-d8-THC , and the magnitude of tolerance development produced by the metabolite was significantly higher than that by d8-THC. The results indicate that ll-hydroxy-d8-THC has an important role not only in the pharmacological effects but also its tolerance development of d8 - THe. THCs and their metabolites competed to the specific binding of CP-55, 940, an agonist of cannabinoid receptor, to synaptic membrane from bovine cerebral cortex. The Ki value of THCs and their metabolites were closely paralleled to their pharmacological effects in mice. A novel cytochrome P450 (cyp2c29) was purified and identified as a major enzyme responsible for the metabolic activation of d8-THC at the II-position in the mouse liver. cDNA of CYP2C29 was cloned from a mouse cDNA library and its sequence was determined. The oxidation mechanism of THC by cyp2c29 was proposed.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2007년도 Proceedings of The Convention
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• pp.57-64
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• 2007

Metabolic interactions of the three major cannabinoids, ${\Delta}^9$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN) with steroid hormones were investigated. These cannabioids concentration-dependently inhibited $3{\beta}$-hydroxysteroid dehydrogenase and $17{\alpha}$-hydroxylase in rat adrenal and testis microsomes. CBD and CBN were the most potent inhibitors of $3{\beta}$-phydroxysteroid dehydrogenase and progesterone $17{\alpha}$-hydroxylase, respectively, in rat testis microsomes. Three cannabinoids highly attenuated hCG-stimulated testosterone production in rat testicular interstitial cells. These cannabinoids also decreased in levels of mRNA and protein of StAR in the rat testis cells. These results indicate that the cannabinoids could interact with steroid hormones, and exert their modulatory effects on endocrine and testicular functions. Metabolic interaction of a THC metabolite, $7{\beta}$-hydroxy-${\Delta}^8$-THC with steroids is also investigated. Monkey liver microsomes catalyzed the stereoselective oxidation of $7{\beta}$-hydroxy-${\Delta}^8$-THC to 7-oxo-${\Delta}^8$-THC, so-called microsomal alcohol oxygenase (MALCO). The reaction is catalyzed by CYP3A8 in the monkey liver microsomes, and required NADH as well as NADPH as an efficient cofactor, and its activity is stimulated by some steroids such as testosterone and progesterone. Kinetic analyses revealed that MALCO-catalyze reaction showed positive cooperativity. In order to explain the metabolic interaction between the cannabinoid metabolite and testosterone, we propose a novel kinetic model involving at least three binding sites for mechanism of the metabolic interactions.

• 생약학회지
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• 제52권2호
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• pp.69-76
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• 2021

Cannabis sativa is an important industrial plant utilized to produce fiber, oil, and medicinal ingredients. A chemotype of cannabis is divided into "Drug type" with predominance of tetrahydrocannabinolic acid (THCA) and "Fiber type" with cannabidiolic acid (CBDA). To develop molecular markers for the discrimination of these two types, nucleotide sequences of THCA synthase and CBDA synthase as well as their pseudogenes were retrieved from the recently published cannabis genome in chromosome scale. Gene-specific SNPs were discovered by multiple alignment of these sequences, and 2 dominant marker sets from each gene were designed for selective amplification. Our markers successfully identified "Drug type" and "Fiber type" cannabis plants as well as forensic samples including processed materials. Our molecular markers will provide a fast and efficient system for molecular-based identification of the cannabis plant.

• 한국작물학회지
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• 제51권5호
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• pp.483-490
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• 2006

저 마약형 대마 품종 육성에 이용하고자 네덜란드 CPRO에서 도입한 저 마약형 유전자원의 특성을 검정한 결과는 다음과 같다. 1. 도입 유전자원들은 고위도인 네덜란드에서 재배했을 때에 비해 중위도인 우리나라에 재배했을 때 개화소요일수가 짧아 경장이 작았다. CBD 함량은 두 지역간 차이가 다소 있는 경향이었으나 THC 함량은 거의 변하지 않았다. 2. 개화소요일수, 경장, 경직경, 생경중 및 섬유수량 등 대마 생육형질들은 서로 간에 고도로 유의한 정의 상관을 보였고, cannabinolds 함량과의 상관에서는 CBD 함량과는 고도로 유의한 정의 상관을 보였으나 THC와 CBN 함량과는 유의한 상관관계를 보이지 않았다. 3. 공시한 44 도입종과 재래종을 군집분석의 Q 상관으로 도출된 r = 0.80으로 선을 그었을 때 3개 품종군으로 나누어졌으며 품종군에도 속하지 않는 것이 4품종이었다. 4. 분류된 모든 품종군은 개화소요일수와 경장이 짧아 우리나라에서 섬유용 품종으로 적합하지 않았고 품종군으로 분류되지 않은 것들 중 도입종 IH3은 THC 함량이 0.08%로 낮으면서 개화소요일수가 122일로 재래종과 비슷하였다.