€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****VIROLOGY***** Yao XJ Wainberg MA Parniak MA Mechanism of inhibition of HIV-1 infection in vitro by purified extract of Prunella vulgaris. In: Virology (1992 Mar) 187(1):56-62 ISSN: 0042-6822 Crude extracts of four Chinese herbs, Arctium lappa, Astragalus membranaceus, Andrographis paniculata, and Prunella vulgaris, were assessed in several tissue culture lines for anti-HIV activity and for cytotoxicity. One extract, obtained from P. vulgaris, was able to significantly inhibit HIV-1 replication with relatively low cytotoxicity. The active factor was purified using sequential precipitations with ethanol and n-butanol, followed by reverse-phase and gel permeation high-performance liquid chromatographic separations. The active component was anionic with a molecular weight of approximately 10 kDa. The purified extract inhibited HIV-1 replication in the lymphoid cell line MT-4, in the monocytoid cell line U937, and in peripheral blood mononuclear cells at effective concentrations of 6, 30, and 12.5 micrograms/ml, respectively. Pretreatment of uninfected cells with the extract prior to viral exposure did not prevent HIV-1 infection. By contrast, preincubation of HIV-1 with the purified extract dramatically decreased infectiousness. The purified extract was also able to block cell-to- cell transmission of HIV-1, prevented syncytium formation, and interfered with the ability of both HIV-1 and purified gp120 to bind to CD4. PCR analysis confirmed the absence of HIV-1 proviral DNA in cells exposed to virus in the presence of the extract. These results suggest that the purified extract antagonizes HIV-1 infection of susceptible cells by preventing viral attachment to the CD4 receptor. Registry Numbers: EC 2.7.7.- (HIV-1 Reverse Transcriptase) EC 2.7.7.49 (RNA-Directed DNA Polymerase) 30516-87-1 (Zidovudine) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ **ABSTRACTS OF THE GENERAL MEETING OF THE AMERICAN SOCIETY FOR MICROBIOL** John JF Kuk R Rosenthal A Synergistic antiretroviral activities of the herb, Prunella vulgaris, with AZT, ddI, and ddC. In: Abstr Gen Meet Am Soc Microbiol (1994) 94:481 (abstract no. S-27) There is a urgent need for new, safe, and inexpensive therapy for HIV infection. We tested a group of commonly available herbs for their activity in aqueous solution against HIV. In screening tests of four commonly used herbs, Prunella vulgaris (PV) was found to exert the best anti-HIV activity. The antiretroviral activity of PV was further analyzed using an aqueous extract prepared by boiling 3 gm in 100 ml. water for 45 min. followed by filter sterilization. The ability of different dilutions of PV to inhibit replication of HIV strain H9/3B was monitored by inhibition of HIV-induced cytotoxicity in MT2 cells, measured by the MTT uptake assay. HIV inhibition of PV alone was tested in four dilutions (undiluted, 1:10, 1:100, and 1:1000). MT2 cell cytotoxicity was present at undiluted and 1:10 dilutions. The 1:100 and 1:1000 dilutions gave 80-90% and 0-25% protection respectively. PV was next tested in combination with AZT, ddI, or ddC. The combinations of PV (1:1000) plus AZT (0.003-0.01 microM) and PV plus ddI (0.1-0.3 microM) gave protection of 69-74% compared to a much lower protection range (10%) by AZT or ddI alone at the same concentrations. Only a slight additive effect was observed for PV plus ddC. We conclude that certain herbs like PV may display synergistic anti-HIV activity when used in combination with antiretroviral nucleosides. Registry Numbers: 30516-87-1 (Zidovudine) 69655-05-6 (Didanosine) 7481-89-2 (Zalcitabine) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****ANTIVIRAL RESEARCH***** Tabba HD Chang RS Smith KM Isolation, purification, and partial characterization of prunellin, an anti-HIV component from aqueous extracts of Prunella vulgaris. In: Antiviral Res (1989 Jun-Jul) 11(5-6):263-73 ISSN: 0166-3542 Prunellin, an anti-HIV active compound, was isolated from aqueous extracts of the Chinese medicinal herb, Prunella vulgaris, and purified to chromatographic homogeneity. Infrared and NMR spectroscopy identified prunellin as a polysaccharide. Elemental analyses, precipitation with calcium(II), barium(II), or 9- aminoacridine suggest a sulfated polysaccharide. Paper chromatography of the exhaustively hydrolyzed material indicates the presence of glucose, galactose, xylose, gluconic acid, galactonic acid and galactosamine as the constituent monosaccharides. The molecular size of prunellin, as determined by gel permeation chromatography and the Squire method on Sephadex G-75, is about 10 kDa. Registry Numbers: 123138-82-9 (prunellin) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ ***CHUNG HSI I CHIEH HO TSA CHIH CHINESE JOURNAL OF MODERN DEVELOPMENTS** Zheng MS Zhang YZ [Anti-HBsAg herbs employing ELISA technique] In: Chung Hsi I Chieh Ho Tsa Chih (1990 Sep) 10(9):560-2, 518 ISSN: 0254-9034 (Published in Chinese) With the aid of the ELISA system this schema represented a laboratory approach to the recognition of anti-HBsAg capability of herbs by using 300 herbal extracts. Altogether 10 herbs (3.0%) were identified as effective. When forming a multiplex plan by employing 10 average P/N ratios as exemplified by 5 varying doses of herb (0.3, 0.6, 1.2, 2.5, 5.0 mg/100 microliters), 2 varying concentrations of HBsAg (10.92, 14.26 P/N ratio), and 3 varying contact time periods (immediate, 1h, 2h) for the comprehensive appraisal of herb efficacy index, these 10 effective herbs were listed in the following order: Prunella vulgaris (1.00), Litchi chinensis (1.26), Gossypium herbaceum (1.45), Cudrania cochinochinensis (1.56), Caesalpinia sappan (1.73), Oldenlandia tenelliflora (1.77), Cautis parthenocissus (1.99), Evodia rutaecarpa (2.01), Portulaca grandiflora (2.44), and Anemone hupehensis (2.83). €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ Zheng M [Experimental study of 472 herbs with antiviral action against the herpes simplex virus] In: Chung Hsi I Chieh Ho Tsa Chih (1990 Jan) 10(1):39-41, 6 ISSN: 0254-9034 (Published in Chinese) Using tissue culture method, the present work with its first-hand observation was primarily concerned with evaluating the antiviral effect of 472 traditional medicinal herbs (comprising raw material 10 mg/ml), through both initial (qualitative) and repeated (quantitative) screens, on type 1 herpes simplex virus. When dealing with water and alcoholic extracts, the effective herbs during initial screens were reduced after repeated screens by a range of 28.8-80.0%. Employing the basic value attained by the simultaneous route of drug administration, a stepwise declining of effective herbs would be: extratube route greater than simultaneous route greater than therapeutic route greater than preventive route. The more the routes of drug administration, the less the multiple-route simultaneous efficacy of a herb. Through repeated screens, 10 highly effective herbs were Aristolochia debilis, Artemisia anomala, Lindera strychnifolia, Patrinia villosa, Pinus massoniana, Prunella vulgaris, Pyrrosia lingua, Rhus chinensis, Sargussum fusiforme and Taraxacum mongolicum. Clinically, among the 78 cases of herpetic keratitis due to HSV1 treated by Pyrrosia lingua and Prunella vulgaris eye drops, a cure was effected in 38 and an improvement in 37, with 3 being of no benefit. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ **CHUNG-KUO CHUNG YAO TSA CHIH CHINA JOURNAL OF CHINESE MATERIA MEDICA* Wang HB Zhang ZY Zia ZX Su Z Li CK [Qualitative analysis of Chinese drug xiakucao (Prunella)] In: Chung Kuo Chung Yao Tsa Chih (1993 Nov) 18(11):655-7, 702 ISSN: 1001-5302 (Published in Chinese) A high performance gas chromatographic procedure has been developed for the quantitation of ursolic acid and oleanolic acid in different parts of Xiakucao collected in different periods. The study provides a scientific foundation for increased availability and rational collecting periods of Xiakucao. Registry Numbers: 508-02-1 (Oleanolic Acid) 77-52-1 (ursolic acid) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****INTERNATIONAL CONFERENCE ON AIDS***** Yamasaki K Nakano M Otake T Kawahata T Mori H Morimoto M Ueba N Anti-HIV-1 activity of Labiatae plants, especially aromatic plants. In: Int Conf AIDS (1996 Jul 7-12) 11(1):65 (abstract no. Mo.A.1062) Object: We have screened and identified compounds that inhibit the replication of HIV-1 from natural products and defined the mechanisms of the anti-HIV-1 activity of the hit, we found some Labiatae plants exhibiting anti-HIV activity, we screened several aromatic plants(herbs) belonging to Labiatae, and searched for the active substances. Method: About 40 herbs belonging to Labiatae used in Japan were extracted in the order acetone, ethanol, 70% ethanol and water. Each extract was investigated for inhibition of the replication of HIV-1(LAV-1). Results and Discussion: Acetone and ethanol extracts of all the samples showed no activity, but the 70% ethanol extract showed light activity. The water extract showed most of the anti-HIV-1 activity. Especially, Prunella vulgaris, Ocimum basilicum L. cv cinnamon, Melissa officinalis L. and Perilla frutescens Britt.var.acta Kudo showed very potent activity at below the concentration of 16 mg/ml. By use column chromatography, we are seaching for common anti-HIV-1 active substances in these herbs. Replication of the fresh isolated HIV-1 strain was inhibited by some extracts. The extracts weakly inhibited the reverse transcriptase activity of HIV-1. All the extracts suppressed giant cell formation in co-cultures of HIV-1-infected and non-infected Molt-4 cells, suggesting that these compounds can inhibit virus adsorption or fusion. Conclusion: Through our study, we found many herbs with anti- HIV-1 activity in in vitro. It is very meaningful that herbs belonging to Labiatae plant, reasonable and easy to cultivate, show very potent anti-HIV-1 activity. It is very important to search for the common active substance in these Labiatae plants. Registry Numbers: EC 2.7.7.- (HIV-1 Reverse Transcriptase) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****MUTAGENESIS***** Horikawa K Mohri T Tanaka Y Tokiwa H Moderate inhibition of mutagenicity and carcinogenicity of benzo[a]pyrene, 1,6-dinitropyrene and 3,9-dinitrofluoranthene by Chinese medicinal herbs. In: Mutagenesis (1994 Nov) 9(6):523-6 ISSN: 0267-8357 The activity of six Chinese medicinal herbs against the environmental mutagens and carcinogens benzo[a]pyrene (B[a]P), 1,6-dinitropyrene (1,6-diNP) and 3,9-dinitrofluoranthene (3,9-diNF) was determined. Samples of Prunella spica, Rheum palmatum, Polygonum multiflorum, Agrimonia pilosa, Ephedra sinica and Teitoutou were tested in an in vitro system. Antimutagenic activity against B[a]P was marked in the presence of extracts (boiled for 2 h in a water bath) whereas that against 1,6-diNP and 3,9-diNF varied from 20 to 86%. The differences in inhibition might be due to inactivation of metabolic enzymes. An extract of P. multiflorum was divided into ether, ethyl acetate and water soluble fractions, which were tested for antimutagenic activity against B[a]P. The antimutagenic action of the ethyl acetate soluble fraction was substantial and dose-dependent. Tannins and related compounds were the major components of the extract, of which epigallocatechin, epigallocatechin gallate, epicatechin gallate and tannic acid strongly inhibited the mutagenicity of B[a]P (2.5 micrograms/plate) in Salmonella typhimurium TA98 with S9 mix. To confirm the results of the in vitro test system, F344/DuCrj male rats were given a subcutaneous injection of B[a]P. Thereafter, they received water extracts of the six Chinese medicinal herbs for 50 weeks and were examined for tumors. The P. multiflorum extract significantly reduced the tumor incidence. Registry Numbers: 22506-53-2 (3,9-dinitrofluoranthene) 42397-64-8 (1,6-dinitropyrene) 50-32-8 (Benzo(a)pyrene) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****MUTATION RESEARCH***** Lee H Lin JY Antimutagenic activity of extracts from anticancer drugs in Chinese medicine. In: Mutat Res (1988 Feb) 204(2):229-34 ISSN: 0027-5107 The antimutagenic activities of extracts of 36 commonly used anticancer crude drugs from Chinese herbs were studied by using the Salmonella/microsomal system in the presence of picrolonic acid or benzo[a]pyrene to test whether they contain direct or indirect antimutagens. Each crude drug was extracted with boiling water for 2 h, the method which is commonly used by Chinese people to prepare the drug for oral intake. The extracts of Pteris multifida P. showed the highest antimutagenic activity against picrolonic acid-induced mutation. The extracts of 6 other different kinds of Chinese herbs were shown to have a moderate antimutagenic activity against picrolonic acid-induced mutation, and they are: Actinidia chinensis P., Artemisia lavendulaefolia DC. and Crotalaria sessiflora L., Prunella vulgaris L., Paris polyphylla S. and Ampelopsis brevipedunculata T. The extracts of Smilax china L., Prunella vulgaris L. and Actinidia chinensis P. were demonstrated to inhibit the mutagenicity of benzo[a]pyrene completely. The 12 other kinds of extracts of Chinese herbs which had a moderate antimutagenic activity against benzo[a]pyrene were: Pteris polyphylla S., Ampelopsis brevipedunculata T., Duchesnea indica F., Gossypium herbaceum L., Lithospermum erythrorrhizon SZ., Artemisia lavendulaefolia DC., Selaginella doederleinii H., Dianthus superbus L., Centipeda minima ABA., Curcuma zedoaria R., Marsdenia tenacissima WA. and Kalopanax septemlobus K. Among them, there were 5 kinds of crude drugs, Actinidia chinensis P., Artemisia lavendulaefolia DC., Prunella vulgaris L., Paris polyphylla S. and Ampelopsis brevipedunculata T., containing antimutagenic factors against both picrolonic acid- and benzo[a]pyrene-induced mutation. Registry Numbers: 50-32-8 (Benzo(a)pyrene) 550-74-3 (picrolonic acid) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****PHARMACEUTICA ACTA HELVETIAE***** Lamaison JL Petitjean-Freytet C Carnat A [Medicinal Lamiaceae with antioxidant properties, a potential source of rosmarinic acid] Lamiacees medicinales a proprietes antioxydantes, sources potentielles d'acide rosmarinique. In: Pharm Acta Helv (1991) 66(7):185-8 ISSN: 0031-6865 (Published in French) Hydroalcholic extracts from four native medicinal Lamiaceae, Lycopus europaeus L., Melissa officinalis L., Origanum vulgare L. and Prunella vulgaris L. have shown significant antioxidative activities, by free radical scavenger effect on DPPH, compared with those of Rosmarinus officinalis L. and Salvia officinalis L. extracts. The antioxidative activity was partly in relation to the rosmarinic acid content. The major hydroxycinnamic compound, quantitatively determinated by HPLC, was present in large amount. The content in Prunella vulgaris L. spikes average 6.1%, based on dry weight. Registry Numbers: 537-15-5 (rosmarinic acid) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****YAKUGAKU ZASSHI. JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN***** Yamasaki K Otake T Mori H Morimoto M Ueba N Kurokawa Y Shiota K Yuge T [Screening test of crude drug extract on anti-HIV activity] In: Yakugaku Zasshi (1993 Nov) 113(11):818-24 ISSN: 0031-6903 (Published in Japanese) The anti-HIV-1 effects of 204 crude drugs of common use in Japan were evaluated in vitro. As a result, 45 samples inhibited HIV-1-induced cytopathogenicity in MT-4 cells. In particular, the hot water extracts of Lithospermum erythrorhizon (root) and Prunella vulgaris (spike) showed the strongest anti-HIV-1 activities. Their IC100 values were both 16 micrograms/ml. In general, the hot water extracts of the crude drug suppressed the replication of HIV-1 growth more strongly than the cold water extracts. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€