Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or... – Inactivation or attenuation; producing viral subunits
Reexamination Certificate
2001-01-18
2002-09-03
Lankford, Jr., Leon B. (Department: 1651)
Chemistry: molecular biology and microbiology
Virus or bacteriophage, except for viral vector or...
Inactivation or attenuation; producing viral subunits
C424S725000, C424S764000, C424S408000, C504S118000
Reexamination Certificate
active
06444458
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to the use of
Tagetes minuta
oil and its components as antiviral agent. More particularly this invention relates to identification of antiviral activity of
Tagetes minuta
oil. This invention also relates to the use of the compounds Z-&bgr;-ocimene and dihydrotagetone present in
Tagetes minuta
oil which are now found to inhibit Carnation Ring Spot (CaRSV) and Carnation Vein Mottle Viruses (CaVMV).
BACKGROUND OF THE INVENTION
Carnation Ring Spot (CaRSV) and Carnation Vein Mottle Viruses (CaVMV) are widespread in carnations and cause appreciable amount of damage. Production of disease free plants and chemical control of vectors are the methods employed for reducing disease incidence in the field (Matthews R. E. F. 1991, Plant Virology, Academic Press, San Diego, pp. 835).
Carnation Vein Mottle virus (CaVMV) is a member of potyvirus group, first reported from U.S.A. and is found all over the world (Kissanis B., 1954, Nature 173:1097). On natural hosts chlorotic and darker green spots, flecks and mottling, flower breaking and malformation symptoms are developed after infection. Virus is transmitted mechanically, and also by aphid vectors.
Chenopodium amranticolor, Chenopodium quinoa
and
Silene pendule
are diagnostically susceptible hosts. On
Chenopodium amaranticolor
chlorotic and necrotic local lesions appear whereas, in
Chenopodium quinoa
, chlorotic lesions with systemic develop after inoculation.
Chenopodium quinoa
and
Dianthus barbatus
are the maintenance and propagation host. It has RNA genone, which is single stranded and virions found in all parts of the host plant. (Morgan J. R., Verhoyen M. and Caneghem, G. V., 1996, Carnation Vein Mottle Potyvirus, In-Viruses of plants, Description and lists from VIDE database ed. Brunt A. A., Crabtree K., Dallwitz M. J., Gibbs A. J. and Watson L. CAB International pp 309-312).
Carnation Ring Spot Virus (CaRSV) first isolated from Dianthus species from U.K. by Kassanis belongs to dianthovirus group (Kassanis B., 1995, Ann. Appl. Biol. 43:103). CaRSV is found all over the world wherever carnations are grown. Virus is transmitted mechanically, and by grafting. Vector transmission is by nematodes. The virus causes leaf mottling, ring spotting, plant stunting, distortion, and flower distortion in host plants whereas, in experimentally infected plants chlorotic and necrotic local lesions, rings and flecks and occasional systemic symptom also appear.
Chenopodium amaranticolor, Chenopodium quinoa
and
Vigna unguiculata
are local lesion assay hosts. Virus can be maintained on
Dianthus barbatus, Nicotiana clevelandii
and
Phaseolus vulgaris
. Virions are isometric, non-enveloped 34 nm in diameter. Gonome of virus consists of RNA, linear, single stranded. Virions are found in all part of the host plant. (Termaine J. H. and Moran J. R., 1996, Carnation Ring spot virus. In-Viruses of plants, Description and lists for VIDE database ed. Brunt A. A., Crabtree K., Dallwitz M. J., Gibbs A. J., and Watson L. CAB International pp 309-312).
The plant volatile oils have been recognised since antiquity to possess biological activity and a number of plant extracts and pure isolates have been mentioned as containing substances which interfere with or inhibit infection of viruses. Some of the compounds like galangin when used in concentrations ranging form 12-47 &mgr;g/ml showed significant antiviral activity against HSV-
1
and Cox B
1
(Meger J. J. M., Afoloyan A. J., a Taylor M. B., Erasmus D., 1997, Antiviral activity of galangin isolated from the aerial parts of
Helichrysum aureonitens, J. Ethnopharma
, 56:165).
Plant Hyptianthera stricta
L. is used against Encephalitis causing viruses pronounced inhibiting activity 75% and 50% CPE inhibition at 62.5 &mgr;g/ml and 15.6 &mgr;g/ml against these two viruses (Saxena G., Gupta P., Chandra K., Lakshmi V., 1997, Antiviral activity of
Hyptianthera shivta
L. against encephalitis causing viruses, Indian Drugs, 34:694). The essential oil of
Melaleuca alternifolia
in concentration of 100, 250, 500 ppm was found to be effective in decreasing local lesions of TMV on host plant
Nicotiana glutinosa
(Bishop C. D., 1995, Antiviral activity of the essential oil of
Melaleuca alternifolia
(Maiden & Betche) cheel (Teatree) against Tobacco Mosaic Virus,. J. Essen. Oil Res 7:641). The chemical composition of essential oil for
Senecio graveleopeus
was analysed by GLC-MS and different components like isovaleraldehyde, &agr;-pinene, sabinene, p-cymene, terpinen-4-ol, &agr; and &bgr;-eudesmone were identified and found to have antimicrobial activity against
Microccus letus, staphylococus aureus
and antifungal activity against
canidida albicans
. The MIC (Minimum Inhibitory Concentration) was 8.73, 10.91 and 2.13×10
−2
mg/ml respectively against all the three organisms. A number of compounds from different plant extracts have antiviral activity (Perez C., Agnese A. M. Cabrere J. L., 1999, The essential oil of
Senecio graveoleus
(Compositae): chemical composition and antimicrobial activity test 66:91). A new acelycated flavonol glycoside, quercetin exhibited IC
50
values of 18.1±1.3 &mgr;g/ml against HIV integrase (Kim J. H., Woo E. R., Shin C. G., Park 1998
, Acer okamotoanum
and its inhibitory activity against HIV-1 integrase. J. Natural Products 61:145). Three new triterpene lactones lancilactones A, B, C together with the known Kadsulactone A were isolated from stems and roots of
Kadsura lancilimba
. Their structure with sterochemistries was determined- from mass and NMR. Compound 3 inhibited HIV replication with an EC
50
value of 1.4 mg/ml and a therapeutic index of greater than 71.4 (Chen F. D., Zhang X. S., Wang K. H., Zhang Y. S., Sun Q. Z., Cosentine L. M., and Lee K. H. 1999, Novel Anti-HIV Lancilactone C and related Triterpense from
Kadsura lancilimba
J. Natural Products 62:94).
Salvia fructiosa
essential oil extracted form aerial parts was analysed by GC-MS. It contained 1,8 cineole (eucalyptol) (47.48%) &agr;+&bgr; thujone (11.93%) and camphor (9.04%). The essential oil of
S. fuctiosa
and its isolated components; thujone and 1,8 cineole, exhibited activity against 8 bacterial strains. Camphor was almost inactive against 8 bacterial strain. Camphor was almost inactive against all the bacteria tested. The essential oil was almost inactive against all the bacteria tested. The essential oil was bactericidal at 1/4000 dilution; dilutions of upto /1/0000 decreased bacterial growth rates. The essential oil of
S. fructicosa
and its three main components exhibited cytotoxic activity against African Green Monkey Kidney (vero) cells and high levels of virucidal activity against Herpes Simplex Virus 1 (Sivropou A., Nikolaou K. E., Kokkini S. L. and Arsenalics M., 1997, Antimicrobial, cytotoxic and antiviral activities of
Saliva fructicosa
essential oil. Journal of Agriculture and Food chemistry, 45: 3197.) The essential oils and their components exhibited inhibiting properties against viruses (Deans S. G. and Waterman P. G., 1993, Biological activity of volatile oils, in: Volatile oil crops, Hay R. K. M. and Waterman P. G. Longman Scientific and Technical pp. 97) fungi (Baruah P., Sharma R. K. Singh, R. S. and Ghosh A. C. 1996, Fungicides activity. of some naturally occurring essential oils against
Fusarium monitiform
, Journal of Essen. Oil Res 8:411) bacteria (Chalchal J. C., Garry R. P., Menut C., Lamaty Li., Malhuret R. and Chopineau, J., 1997, correlation between chemical composition and antimicrobial activity VI, Activity of some African essential oils, Journal of Essen. Oil Res. 9:67) malaria (Milnau G., Valentin A., Benoit R., Mallie M., Bastide J. M., 1997, in vitro antimalarial activity of eight essential oils, Journal of Essen. Oil Res. 9:329). These are very few reports on effects of essential oils on viruses or viral infection in either animals or plants.
Tagetes minuta
L. (Asteraceae) grows wild and yields essential oil having commercial value in perfumery and flavour industry (Handa K. L.,
Joshi Virendara Prasad
Ram Raja
Sharma Anupama
Singh Bikram
Zaidi Aijaz Asghar
Council of Scientific and Industrial Research
Ladas & Parry
Lankford , Jr. Leon B.
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