IYER PRIYA R (IN)
IYER PRIYA R (IN)
CHRISTINA L. NANCE ET AL.: "Preclinical development of the green tea catechin, epigallocatechin gallate, as an HIV-1 therapy", JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, vol. 123, no. 2, pages 459 - 465, XP025907166
FAROOQ, M.U. ET AL.: "Gold Nanoparticles-enabled Efficient Dual Delivery of Anticancer Therapeutics to HeLa Cells", SCIENTIFIC REPORTS, vol. 8, no. 2907, 13 February 2018 (2018-02-13), XP055744441
| I CLAIM: 1. A biosynthesized nano drug, comprising of Camellia sinensis extract as a reducing agent and gold salt, wherein said biosynthesized nano drug posses anti HIV activity. 2. The biosynthesized nano drug as claimed in claim 1, wherein part of the Camellia sinensis plant used is leaves. 3. The biosynthesized nano drug as claimed in claim 1, wherein the gold salt is chloroauric acid (HAUC14). 4. The biosynthesized nano drug as claimed in claims 2 and 3, wherein the chloroauric acid and leaf extract of Camellia sinensis plant are in the ratio of 10: 1. 5. The biosynthesized nano drug as claimed in claim 4, wherein gold nanoparticles synthesized at 10 concentration posses anti HIV activity. 6. The biosynthesized nano drug as claimed in claim 5, wherein the synthesized Camellia sinensis (CS) mediated gold nanoparticles - CS AuNPs has 12.84% of gold and 87.16% of leaf extract of Camellia sinensis plant. 7. The biosynthesized nano drug as claimed in claim 6, wherein the synthesized Camellia Sinensis (CS) mediated gold nanoparticles - CS AuNPs has EC50 value of 13.26 ng/ml and percentage of inhibition is 99.984%. 8. The biosynthesized nano drug as claimed in claim 6, wherein the gold nanoparticles is in the size range of about 15 - 30 nm in diameter. 1 |
FIELD OF INVENTION
The present invention relates to a drug for anti HIV activity, particularly to bio synthesized nano drug for treatment of HIV/AIDS.
BACKGROUND OF INVENTION
Nanotechnology is the study and application of extremely small things and can be used across all the science fields. Because of the nanoparticle's size, catalytic property, ability to deliver drug, increased efficacy, and decreased toxicity, nanotechnology have become significant in many fields in the recent years, such as but not limited to energy, health care, environment, and agriculture. Conventionally, nanoparticles have been synthesized by various physical and chemical methods, having negative impact on environment. The production of nanoparticles from environmentally friendly green biological methods is alternative to the conventional methods. Biosynthesis of nanoparticles using leaves extracts and fruit extracts have been disclosed in prior arts. The synthesis of nanoparticles using plant extracts is gaining importance due to the various advantages like green chemistry, low cost and eco safe. Chemical drugs are being used for the HIV/AIDS treatment, which is expensive but results in various side effects.
Various plants are listed in literature which on research found to have anti cancer properties and anti oxidant properties when used as crude extract. But however, bio synthesized nano formulations with high anti HIV activity that could be used in place of existing chemical drugs or in combination with the current drugs does not exist.
US20080199488 discloses an anti-AIDS agent which comprises, as the active ingredients, one or more components selected from the group consisting of MelastomavillosumLodd., DipterocarpusobtusifoliusTeijsm ex Miq., Lyophyllumaggregatum, Dictyophoraindusiata, pu-erh tea, mentha and stevia. However, Pu-erh tea is a type of fermented green tea, the production method thereof comprising fermentation of green tea with Aspergillus for more than three years.
CN 1621084 relates to one anti-HIV product containing active green matter, and the anti-HIV product may be used as natural medicine in various forms, health foods in various forms and food additive. The anti-HIV product contains green tea, natural anti-HIV medicine and/or food, and has outstanding anti -HIV effect, low cost and no toxic side effect. However, in the cited document the green tea extract alone and in combination with other foods is used as a supplement. Moreover, the quantity of green tea used is 20 ~ 30g, per day.
JP2002518454 relates to herbal extract composition comprising extract of four ingredients namely ARUM, extract of POMEGRANATE, extract of HIBISCUS and extract of TEA. The herbal extract composition of the invention demonstrates in vitro stimulation of lymphocyte transformation and cytokine production, and in vitro inhibition of gpl20 binding, and provides a potential candidate for therapies and treatments for immune disorders and HIV infection.
Non Patent literature- Gold Bull (2012) 45:53-59 DOI 10.1007/sl3404- 012-0048-7 talks about the combination effects of trivalent gold ions and gold nanoparticles with different antibiotics against resistant Pseudomonas Aeruginosa. The combination effect of gold materials including trivalent gold ions (Au3+) and gold nanoparticles (Au NPs) with 14 different antibiotics was investigated against the clinical isolates of P. aeruginosa, Staphylococcus aureus and Escherichia coli.
Accordingly, there exists a need for bio synthesized nano drug for treatment of HIV/AIDS.
OBJECTS OF INVENTION
One or more of the problems of the conventional prior art may be overcome by various embodiments of the present invention.
It is the primary object of the present invention to provide nano formulation for treatment of HIV/AIDS.
It is another object of the present invention to provide bio synthesized nano drug for treatment of HIV. It is another object of the present invention, wherein the bio synthesized nano drug could be used in combination with existing drugs, so as to enhance its efficiency and as well to reduce the side effects over long term usage of chemical drugs.
It is another object of the present invention to provide bio synthesized nano drug which eliminates the use of toxic chemicals.
Another object of the present invention is to provide bio synthesized nano drug comprising of Chloroauric acid and Camellia sinensis extract, said Camellia sinensis extract is used as a reducing agent.
It is another object of the present invention, wherein the Chloroauric acid and Camellia sinensis extract are in the ratio of 10: 1.
It is another object of the present invention, wherein the Chloroauric acid is in about 10 concentration at pH 8.
SUMMARY OF INVENTION
Thus, according to the basic aspect of the present invention there is provided a biosynthesized nano drug, comprising of Camellia sinensis extract as a reducing agent and gold salt, wherein said biosynthesized nano drug posses anti HIV activity.
It is another aspect of the present invention, wherein part of the Camellia sinensis plant used is leaves.
It is another aspect of the present invention, wherein the gold salt is chloroauric acid (HAuCL t ).
It is another aspect of the present invention, wherein the chloroauric acid and leaf extract of Camellia sinensis plant are in the ratio of 10: 1. It is another aspect of the present invention, wherein gold nanoparticles synthesized at 10 concentration posses anti HIV activity.
It is another aspect of the present invention, wherein the synthesized Camellia Sinensis (CS) mediated gold nanoparticles - CS AuNPs has 12.84% of gold and 87.16% of leaf extract of Camellia sinensis plant.
It is another aspect of the present invention, wherein the synthesized Camellia Sinensis (CS) mediated gold nanoparticles - CS AuNPs has EC 50 value of 13.26 ng/ml and percentage of inhibition is 99.984%.
It is another aspect of the present invention, wherein the gold nanoparticles is in the size range of about 15 - 30 nm in diameter.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1: illustrates screening of the nanocompounds for their anti viral properties against HIV according to the present invention.
Figure 2: illustrates synthesized Camellia Sinensis (CS) mediated gold nanoparticles - CS AuNPs according to the present invention.
Figure 3: is a graphical representation of concentration of nanocompounds vs percentage of inhibition to show the anti HIV activity according to the present invention.
DETAILED DESCRIPTION OF THE INVENTION WITH REFERENCE TO ACCOMPANYING DRAWINGS
The present invention as herein described relates to bio synthesized nano drug for treatment of HIV, said drug could be used in combination with existing drugs, so as to enhance its efficiency and as well to reduce the side effects over long term usage of chemical drugs. The bio synthesized nano drug comprises of Chloroauric acid and Camellia sinensis extract, said Camellia sinensis extract is used as a reducing agent. The Camelia sinensis - green tea extract is used for the synthesis of a nano formulation. As used herein, the term“nanoparticle” refers to a particle having at least one dimension and sized between 1 and 100 nanometers. The following examples will further illustrate the process of preparing gold nanoparticles from Camelia sinensis - green tea extract.
Example 1:
Preparation of the Camelia sinensis - green tea extract:
About 5g of the Camelia sinensis leaves were washed well and taken in a conical flask, and boiled for 5-10 minutes with about 50ml water to form extract. The extract was allowed to cool, filtered and collected through Whatman filter paper. The filtrate or the filtered extract were stored at 4° C and used for further synthesis. The Camelia sinensis - green tea extract is used for the synthesis of a nano formulation such that, the green tea’s bio active components were brought within a gold nanoparticle.
Example 2:
Preparation of the stock solution-Chloroauric acid:
About 0.03g of Chloroauric acid (HAuCU) is mixed in about 1000ml of double distilled water and dissolved well, said dissolved solution with concentration of 10 M is used as stock solution for further experiments. The pH is adjusted to alkaline ranging from pH 7 to pH 8.
Example 3:
Synthesis of gold nanoparticles:
For the synthesis of gold nanoparticles (AuNPs), about 10 ml of the stock solution were taken and adjusted for alkaline pH ranging from pH 7 to pH 8. To this, 1 ml of the plant extract were added, thereby the ratio of 10: 1 is followed as the constant ratio for production of larger quantities. The green tea extract is used as the reducing agent for the synthesis. The nanoparticles synthesis was completed in about 10 minutes which was visibly seen from the color change of the stock solution from pale yellow to deep wine red. The bio synthesized nanoparticles was used as a drug for the treatment of HIV/AIDS.
In an aspect, the size of bio synthesized nano particles ranges between 15 - 30 nm in diameter and is spherical in morphology. Table 1 shows summary of gold nanoparticle synthesis by varying chloroauric acid solution concentration.
Table 1 :
The concentration of the precursor solution was prepared in different concentrations and was used for the synthesis as shown in Table 1. The gold nanoparticles were successfully synthesized in all the concentrations. Upon screening for anti HIV activity, the concentration 10 proved to have significant anti HIV activities.
Table 2 shows summary of gold nanoparticle synthesis by varying Camelia sinensis leaf extract concentration. Table 2:
The synthesis of gold nanocompounds was carried out in different ratios of precursor solution: plant extract as shown in Table 2. The gold nanoparticles were successfully synthesized in all the various ratios under study. Upon screening for anti HIV activity, the ratio of 10: 1 of chloroauric acid: plant extract proved to have significant anti HIV activities.
Example 4:
Anti HIV activity study:
Testing:
The in vitro testing was carried out to check the anti HIV activity, which showed an Inhibition percentage at 99.98% at the Concentration of 19.5ng/ml for the HIV virus around for the treated compared to the untreated. ICP - OES ANALYSIS (Inductively coupled plasma optical emission spectrometry)
PERKIN ELMER OPTIMA 5300 DV ICP-OES Table 3:
On analysis, the above result as shown in Table 3 indicates the concentration of the metallic component present in the gold nanocompounds. This implies that the synthesized nanocompounds are made up of 12.84% of the elemental substance gold and 87.16% of medicinal plant extracts.
Screening of the nanocompounds for their anti viral properties against HIV :
Referring to Figure 1, TZM-bl cells were placed in a 96-well format (1 x 10 4 cells/well) for 24 hrs before infection. The viral supernatant with 0.005 MOI was pre-incubated with different dilutions of the nanocompounds (10-fold dilutions starting from 1x10 ng/ml to 1x10 ng/ml) for 2 hrs at 37°C before infection to the TZM-bl cells. At 40 to 48 hrs post infection, cells were lysed with IX passive lysis buffer and luciferase expression was measured as relative light units using the Bright-Glo luciferase assay system. The EC50 value of each nanocompound was calculated using the Graph pad prism5 software. Table 4 shows the EC50 value of gold nanoparticle which is 13.26 ng/ml and percentage of inhibition is 99.984%. Figure 2 illustrates synthesized Camellia Sinensis (CS) mediated gold nanoparticles - CS AuNPs
Table 4:
RESULTS:
The anti HIV activity was checked for gold nanoparticles synthesized at different concentrations at 10 -1 , 10 -2 and 10 -3 and at various ratio of Chloroauric acid: Plant extract. Among all these tested gold nanoparticles, the gold nanoparticles synthesized at 10 concentration in 10: 1 ratio gave the successful anti HIV activity as shown in Table 5 and Figure 3.
Table 5: