Indian Patents. 269768:“A METHOD OF MAKING AN ARSENIC FREE FORMULATION FOR SUVARNA BHASMA”

Title of Invention	“A METHOD OF MAKING AN ARSENIC FREE FORMULATION FOR SUVARNA BHASMA”
Abstract	An arsenic free safe suvarna (gold) bhasma substitute consisting of biologically stabilized gold nano particles .

Full Text	FORM - 2 THE PATENTS ACT, 1970 (39 OF 1970) AND THE PATENTS RULES, 2003 COMPLETE SPECIFICATION (See section 10; rule 13} A FORMULATION WHICH IS SAFE SUBSTITUTE FOR GOLD BHASMA NANO CUTTING EDGE TECHNOLOGY PVT. LTD. an Indian Company of 79/87, D. Lad Path, Kalachowki, Mumbai 400 033, Maharashtra, India THE FOLLOWING SPECIFICATION PARTICULARLY DESCRIBES THE INVENTION AND THE MANNER IN WHICH IT IS TO BE PERFORMED This invention relates to a formulation which is a safe substitute for gold bhasma . What is envisaged in accordance with this invention is a formulation containing gold nano particles stabilized with biological tissue extracts. Gold promotes a general euphoric feeling it helps the body to relax, enhances the body's natural defences against illness and promotes vitality and longevity. It has been found to improve glandular functions. It may raise energy levels and may also have anti-inflammatory effects. Some signs of a deficiency are: 1. Arthritis 2. Brain dysfunction 3. Chills 4. Circulatory disorders 5. Depression 6. Heat flashes 7. Insomnia. Gold has also long been used for medicinal purposes. The first recorded use of gold as a medicine took place in Alexandria, Egypt. A group known as alchemists (the best known of which was Paraclesus) purportedly made a solution of gold water, which had the ability to restore youth and perfect health. In the West, gold was used during the Middle Ages as a heath restorative. But it wasn't until 1857 that gold was prepared in a sufficiently pure state for widespread therapeutic use. Traditionally, gold has been used for a variety of conditions including alcoholism, arthritis, skin ulcers, burns, and various types of punctures and for nerve-end operations. Gold has been medically documented as a highly effective remedy in certain cases of inoperable cancer. Used alternately with silver, gold helps build strong natural defences against disease and promotes renewed vitality and longevity. Today, Gold reported to have a tremendous balancing and harmonizing effect on both physical and emotional states. Jt can be particularly useful in treating those who suffer from unstable mental and emotional states such as depression, Seasonal attitude Disorder (SAD), general states of melancholy, sorrow, fear, despair, anguish, frustration and those with suicidal tendencies. Alcohol Addiction : Gold is also reported to weaken the desire for alcohol and is used in treating alcoholic addiction. Rejuvenating Glands: Gold has been used in cases of glandular and nervous in coordination, to help rejuvenate glands and to stimulate nerves and release nervous pressure. Healing the Heart: Gold is also though to have a direct effect on the rhythmic, balancing, and activity of the heart and for improving blood circulation and rejuvenating sluggish organs, especially the brain. Digestive Help: Gold is beneficial to the digestive system and the body's warmth mechanism is positively affected by gold, particularly in cases of chills, heat flashes and night sweats. Additional reported benefits of Gold : • A natural antibiotic • Stops infections form a burn and • Strengthens the natural immune helps healing without scaring defenses • Improves memory • Eliminates skin ulcers • Enhances libido/sexual; • Helps relieve chills, hot flashes and performance night sweats • Promotes deep ram sleep In the ayurvedic, unani and homeopathid systems of medicine the therapeutic uses of gold in various forms has been well documented. In the homeopathic system of medicine, aurum metallicum has been used for treatment of the bones and faulty degeneration. In this system gold is used in low dilutions for these ailments. In the ayurvedic and unani systems, calx of gold [gold bhasma ] is almost treated as an elixir of life and is considered as a panacea for all ills. There is an elaborate process for the manufacture of gold bhasma [ suvarna bhasma]. Gold bhasma is prepared by rubbing together 2 parts of mercury with one part of gold along with lemon juice to form a mass. This mass is along with 3 parts of sulfur is pounded in a mortar and pestle. The resultant is place in a crucible, covered and heated. There are many variations of this basic process. For example it is common to add substances such as arsenic sulfide and extracts of plants such as horse gram (Dolichos biflorus) and pounding the mixture with gold. The process is repeated up to 14 times till the gold completely loses its metallic character and becomes reduced to a dark powder. It takes almost three months to manufacture a significant quantity of the bhasma and the process of manufacture is immensely complicated. Apart from being time consuming the process is expensive in today's context. In addition, the manufacture involves the use of mercury, arsenic and sulfur which are by themselves considered toxic. The use of gold particles has been suggested in numerous patents. US patent no. 6,530,944 describes the use of Gold Nano Particles for the localized treatment of cancer, inflammation or other disorders involving over proliferation of tissue by locally inducing hyperthermia. US patent no. 6,818,199 describes use of gold nano particles for enhanced medical imaging. US patent no. 6,878,814 describes gold NP nanoparticle-oligonucleotide conjugates for detecting and sequencing nucleic acids for diagnosis of genetic, bacterial, and viral diseases. US patent Application no 20030176335 relates to methods of forming gold nano structures for delivering drugs and other guest compounds across a host membrane and to pharmaceutical compositions comprising such compounds. Recently, Gold nano particles were shown to enhance radiotherapy in mice. Rheumatoid arthritis, commonly referred to as RA, is the second most crippling disease in man, ranking immediately behind cardiovascular defects. Until recently no effective treatment for RA has been known. Gold compounds generally used for treatment of RA and other conditions are Auranofin, Aurothioglucose , Gold Sodium Thiomalate, available as oral capsules, and parenteral injections. However they are known to cause allergies, soreness of tongue, metallic taste, skin-rash gum-bleeding ulcers or white spots on lips or in mouth and throat. Breathlessness, Abdominal pain, cramping, tarry stools, decreased urination has also been reported . US patent 4,315,028 describes a method of treating RA with a complex of penicillamine or N-acetyl penicillamine with gold or copper. However complex will not be useful in patients with penicillamine resistance. US patent 4,405,311 describes a method and apparatus for treating rheumatoid arthritis by direct injection of electrically generated gold ions into the patient's joint. This therapy is expensive, cumbersome and hence difficult to use on a mass scale. US patent 4,606,354 describes an implant for the treatment arthritis which comprises a carbon fiber having a discontinuous coating of gold thereon, exposing the carbon in patches. The carbon and gold form a galvanic couple which is implanted in an arthritic joint and in the presence of body fluids acts like a battery and released gold ions continuously, thus relieving pain. However, the therapy is quite expensive. This invention envisages the use of gold nano particles as a safe substitute for suvarna bhasma having the same if not better properties and which can be made inexpensively very quickly and does not require the use of mercury, arsenic or sulfur in its manufacture. According to this invention there is provided a formulation containing gold nano particles stabilized by an aqueous extract of biological tissue , said particles dispersed in a carrier in which the concentration of the gold nano particles ranges between 10 to 250 ppm. The method of making the biologically stabilized gold nano particles has been described in detail in our copending patent application no: 550/MUM/2004 and involves a process for making gold sub micron scale particles comprising the steps of: (a) dissolving a salt of the gold in water having conductivity less than 3 micro Siemens to obtain a solution in which the concentration of gold ions is in the range of 10, 000 to 25 000 ppm , (b) preparing a fresh filtered aqueous solution of biological tissue extract; (c) diluting the aqueous solution with deionized water in the ratio ranging from 1:1 to 1: 25 to form a solution having a open circuit potential + 0.1 and +0.2 volt and a pH between 5.5 to 7.5 and a total organic carbon content of at least 7,500 ppm; (d)maintaining the said aqueous solution under continuous agitation at a temperature between 20 and 30 degrees Celsius; (e) inoculating a minute quantity of the gold salt solution in the said aqueous extract solution under continuous agitation such that the final concentration of the metal ion in the reaction mixture is in the range of ] 0 to 250 ppm; (f) continuing the agitation for a period of 30 minutes to 5 hours to obtain a colloidal suspension of gold nano particles; (g)separating the nano particles from colloidal suspension by a known process such as centrifugation. The carrier may be a deionized water solution for injection or drops, an aerosol spray for inhalation, a granule, a globule or tableting formulation Typically the biological tissue is derived from macerated plant cells selected from a group of plants consisting of: Alfalfa (Medicago sativa), Blood wort (Achillea millefolium), Garlic (Allium sativum) Indian aloe(Aloe barbadenesis), Celery(Apium graveolens) Ginger(Zingiber officinale), Castor seeds(Ricinus communis) Chirayata(Swertia chirayata), Colchium(Colchicum luteum) Gokulakanta(Hygrophila spinosa), Indian Gooseberry(Emblica officinalis), Indian Sarsaparilla (Hemidesmus indicus), Indian Senna(Cassia augustifolia), Indian Squill (Urginea indica), Lead wort(Plumbago zeylanica), Lemon Grass (Cymbopogon citratus), Madhuca(Madhuca indica), Nutmeg(Myristica fragrans), Pepper(Piper nigrum), Rosemary(Rosamarinus officinalis), Saffron(Crocus sativus) Sage(Salvia officinalis), Turpeth(Operculina turpethum), Vasaka(Adhatoda vasica), Winter cherry(Withania somnifera) Pergularia(Pergularia extensa), Onion(Allium cepa), Coriander(Coriandrum sativum,Linn), Turmeric(Curcuma longa), and Tinospora cordifolia, Jatamansi [nordosstachys jatamansi], shatavari [asparagus racemosus] , triphala, brhami[ bocopa monnieri], Kappikachhu [Mucuna pruriens], deadly nighshade [atropa belladonna], and Centella asiatica. The invention will now be described with reference to the accompanying text and drawings. During the course of this invention ten gold bhasma samples available commercially very chemically analyzed and were found to have average gold concentration in the region of 82%. In addition elements such as iron (2%), sulfur (7%), oxygen (8%), arsenic (0.7%) and mercury (0.3%) were detected. The sizes of gold particles in the bhasma were analyzed by scanning electron microscopy and atomic force microscopy. Figure 1 in the accompanying drawings is a three dimensional atomic force microscope image of gold particles seen in one of the commercially obtained gold bhasma and figure 2 in the accompanying drawings is a gross graphical representation of particle size found in another gold bhasma sample. Figure 3 in the accompanying drawings is a three dimensional AFM image of yet another commercially obtained sample of gold bhasma showing gold particles. It was seen that the sizes of gold particles in the commercially obtained suvarna bhasma ranged between 23 nm and almost 250nm with the majority being above 150 nm. These data indicate that the well documented therapeutic activity exhibited by suvarna bhasma is chiefly because of its gold content in the form sub micron size particles. It was therefore envisaged that if the gold particles in suvarna bhasma are replaced by gold nano particles biologically stabilized and therefore biologically compatible, the resulting formulation will exhibit similar therapeutic activity. To test this hypothesis during the course of present invention a comparative study on in vitro protease inhibition activity of suvarna bhasma and gold nano particles was carried out. Protease inhibition activity was selected because a variety of proteases such as matrix metallo proteinases [MMP], collagenases are overtly implicated in rheumatoid diseases. Therefore, protease inhibition is considered the most important parameter of therapeutic activity during the study. Figure 4 in the accompanying drawings is a table showing the comparative percentage inhibition activity of gold nano particles in accordance with this invention, gold nano particles made conventionally by a chemical capping process and the activity of gold in gold bhasma. As can be seen in the table in figure 4 of the drawings, the protease inhibition activity of the gold nano particles in accordance with this invention is favourably comparable with the activity of gold bhasma and in some cases even significantly better than gold bhasma. A careful selection from the group of plant tissue from the list of plant described above is made and the gold nano particles can be used for specific disorders. For instance, a combination of aqueous extracts of withania somnifera, ginger, turmeric, pepper and tinospora cordifolia may be used in combination as the stabilizing solution for making a drug for the treatment of arthritis. Similarly, a combination of withania somnifera, brahmi, triphala, and centella asiatica may give better results in treating neuro pathological disorders. Thus the gold nano particles biologically stabilized is a safefnon arsenic containing] substitute for gold bhasma with comparable activity. We Claim: [ 1] An arsenic free formulation for suvarna bhasma comprising (.1) biologically stabilized gold nano particles in the size range of 1 to 100 mm; and (2) a carrier iin which the concentration of the said biologically stabllized gold nano particles is in the range of 80%. [2] An arsenic free formulation as claimed in claim 1, in which the gold nano particles are stabilized biologically with an aqueous solution of macerated plant tissue cells . [3] An arsenic free formulation as claimed in claim 2, in which the aqueous solution is diluted in deionised water in the up to ten folds. [4] An arsenic free formulation as claimed in claim 2, in which the plant tissue is at least one plant tissue selected from a group of plant tissues which include leaves, roots, stems, flowers and fruits of the following plants Alfalfa (Medicagosativa), Blood wort (Achillea millefolium), Garlic (Allium sativum) Indian aloe(Aloe barbadenesis), Celery(Apium _ Igraveolens) Ginger(Zingiber officinale), Castor seeds(Ricinus communis) ' Chirayata(Swertia_chirayata)3 Colchium(CoIchicum luteum) Gokulakanta(Hygrophila spinosa), Indian Gooseberry(Emblica officinalis), Indian Sarsaparilla(Hemidesinus Jndicus), Indian Senna(Cassia augustifolia), Indian Squill(Urginea indica), Lead wort(Plumbago zeylanica), Lemon Grass(Cymbopogon citrarus), Madhuca(Madhuca indica), Nutmeg(Myristica fragrans), Pepper(Piper nigrum), Rosemary(Rosamarinus officinalis), Saffron(Crocus sativus) Sage(Salvia officinalis), Turpeth(Operculina turpethum), Vasaka(Adhatoda vasica), Winter cherry(Withania somnifera) Pergularia (Pergularia extensa), Onion(Allium cepa), Coriander(Coriandrum satiyum,Linn), Turmeric(Curcuma longa), and Tinospora cordifolia, Jatamansi [nordosstachys jatamansi"], shatavari La'sparagus racemosus] , triphala, brhami[bocopa monnieri], Kappikachhu [Mucuna pruriens], deadly nighshade [atropa belladonna], and Centella asiatica. [5] An arsenic free formulation as claimed in claim 1 or 2, in which the carrier is a liqujd, suspension, aerosol spray granules, globules, dry powder, and a tableting formulation . [6] A method of making an antimicrobial formulation.aecording to any one of the preceding claims, which includes the steps of (1) making a carrier selected from a group of carriers which includes a liquid, suspension, aerosol spray granules, globules, dry powder, and a tableting formulation in a conventional manner, (2) making an aqueous dispersion of biologically stabilized gold nano particles in the size range of 1 to 100 nm; (3) mixing a dispensed quantity of the said aqueous dispersion in the said carrier to form a homogenous matrix in which the concentration of the biologically stabilized gold nano particles is in the range of 10,000 to 25, 000 ppm. [7] A method of making an arsenic free gold formulation as claimed in claim 6, in which the aqueous dispersion.of biologically stabilized gold nano particles is made by the steps of (a) dissolving a salt of gold in water having conductivity less than 3 micro Siemens to obtain a solution in which the concentration of gold ions is in the range of 50 000 ppm; (b) preparing a fresh filtered aqueous solution of biological tissue extract; (c) diluting the aqueous solution with deionized water upto ten fold to form a solution having an open circuit potential between + 0.2 and +0.2 volt and a pH between 5.5 to 7.5 and total organic carbon content at least 7,500 ppm; (d) maintaining the said aqueous solution under continuous agitation at a temperature between 20 and 30 degrees Celsius; (e) inoculating gold salt solution in the said aqueous extract solution under continuous agitation such that the final concentration of the metal ion in the reaction mixture is in the range of 10 000 to 25 000 ppm; (f) continuing the agitation for a period of 30 minutes to 3 hours ; to; obtain a colloidal suspension of gold nano particles; (g)separating the nano particles from colloidal suspension by a known process such as centrifugation.

Full Text

FORM - 2
THE PATENTS ACT, 1970
(39 OF 1970)
AND
THE PATENTS RULES, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13}
A FORMULATION WHICH IS SAFE SUBSTITUTE FOR
GOLD BHASMA
NANO CUTTING EDGE TECHNOLOGY PVT. LTD.
an Indian Company of 79/87, D. Lad Path, Kalachowki, Mumbai 400 033, Maharashtra,
India
THE FOLLOWING SPECIFICATION PARTICULARLY DESCRIBES THE INVENTION AND THE MANNER IN WHICH IT IS TO BE PERFORMED

This invention relates to a formulation which is a safe substitute for gold bhasma .
What is envisaged in accordance with this invention is a formulation containing gold nano particles stabilized with biological tissue extracts.
Gold promotes a general euphoric feeling it helps the body to relax, enhances the body's natural defences against illness and promotes vitality and longevity. It has been found to improve glandular functions. It may raise energy levels and may also have anti-inflammatory effects. Some signs of a deficiency are:
1. Arthritis
2. Brain dysfunction
3. Chills
4. Circulatory disorders
5. Depression
6. Heat flashes
7. Insomnia.
Gold has also long been used for medicinal purposes.
The first recorded use of gold as a medicine took place in Alexandria, Egypt. A group known as alchemists (the best known of which was Paraclesus) purportedly made a solution of gold water, which had the ability to restore youth and perfect health.
In the West, gold was used during the Middle Ages as a heath restorative. But it wasn't until 1857 that gold was prepared in a sufficiently pure state for widespread therapeutic use. Traditionally, gold has been used for a

variety of conditions including alcoholism, arthritis, skin ulcers, burns, and various types of punctures and for nerve-end operations.
Gold has been medically documented as a highly effective remedy in certain cases of inoperable cancer.
Used alternately with silver, gold helps build strong natural defences against disease and promotes renewed vitality and longevity.
Today, Gold reported to have a tremendous balancing and harmonizing effect on both physical and emotional states. Jt can be particularly useful in treating those who suffer from unstable mental and emotional states such as depression, Seasonal attitude Disorder (SAD), general states of melancholy, sorrow, fear, despair, anguish, frustration and those with suicidal tendencies.
Alcohol Addiction :
Gold is also reported to weaken the desire for alcohol and is used in treating alcoholic addiction.
Rejuvenating Glands:
Gold has been used in cases of glandular and nervous in coordination, to help rejuvenate glands and to stimulate nerves and release nervous pressure.
Healing the Heart:

Gold is also though to have a direct effect on the rhythmic, balancing, and activity of the heart and for improving blood circulation and rejuvenating sluggish organs, especially the brain.
Digestive Help:
Gold is beneficial to the digestive system and the body's warmth mechanism is positively affected by gold, particularly in cases of chills, heat flashes and night sweats.
Additional reported benefits of Gold :
• A natural antibiotic • Stops infections form a burn
and
• Strengthens the natural immune helps healing without scaring defenses • Improves memory
• Eliminates skin ulcers • Enhances libido/sexual;
• Helps relieve chills, hot flashes and performance night sweats
• Promotes deep ram sleep
In the ayurvedic, unani and homeopathid systems of medicine the therapeutic uses of gold in various forms has been well documented.
In the homeopathic system of medicine, aurum metallicum has been used for treatment of the bones and faulty degeneration. In this system gold is used in low dilutions for these ailments.
In the ayurvedic and unani systems, calx of gold [gold bhasma ] is almost treated as an elixir of life and is considered as a panacea for all ills. There is an elaborate process for the manufacture of gold bhasma [ suvarna bhasma]. Gold bhasma is prepared by rubbing together 2 parts of mercury

with one part of gold along with lemon juice to form a mass. This mass is along with 3 parts of sulfur is pounded in a mortar and pestle. The resultant is place in a crucible, covered and heated. There are many variations of this basic process. For example it is common to add substances such as arsenic sulfide and extracts of plants such as horse gram (Dolichos biflorus) and pounding the mixture with gold. The process is repeated up to 14 times till the gold completely loses its metallic character and becomes reduced to a dark powder. It takes almost three months to manufacture a significant quantity of the bhasma and the process of manufacture is immensely complicated. Apart from being time consuming the process is expensive in today's context. In addition, the manufacture involves the use of mercury, arsenic and sulfur which are by themselves considered toxic.
The use of gold particles has been suggested in numerous patents.
US patent no. 6,530,944 describes the use of Gold Nano Particles for the localized treatment of cancer, inflammation or other disorders involving over proliferation of tissue by locally inducing hyperthermia.
US patent no. 6,818,199 describes use of gold nano particles for enhanced medical imaging.
US patent no. 6,878,814 describes gold NP nanoparticle-oligonucleotide conjugates for detecting and sequencing nucleic acids for diagnosis of genetic, bacterial, and viral diseases.
US patent Application no 20030176335 relates to methods of forming gold nano structures for delivering drugs and other guest compounds

across a host membrane and to pharmaceutical compositions comprising such compounds.
Recently, Gold nano particles were shown to enhance radiotherapy in mice.
Rheumatoid arthritis, commonly referred to as RA, is the second most crippling disease in man, ranking immediately behind cardiovascular defects. Until recently no effective treatment for RA has been known.
Gold compounds generally used for treatment of RA and other conditions are Auranofin, Aurothioglucose , Gold Sodium Thiomalate, available as oral capsules, and parenteral injections. However they are known to cause allergies, soreness of tongue, metallic taste, skin-rash gum-bleeding ulcers or white spots on lips or in mouth and throat. Breathlessness, Abdominal pain, cramping, tarry stools, decreased urination has also been reported .
US patent 4,315,028 describes a method of treating RA with a complex of penicillamine or N-acetyl penicillamine with gold or copper. However complex will not be useful in patients with penicillamine resistance.
US patent 4,405,311 describes a method and apparatus for treating rheumatoid arthritis by direct injection of electrically generated gold ions into the patient's joint. This therapy is expensive, cumbersome and hence difficult to use on a mass scale.
US patent 4,606,354 describes an implant for the treatment arthritis which comprises a carbon fiber having a discontinuous coating of gold

thereon, exposing the carbon in patches. The carbon and gold form a galvanic couple which is implanted in an arthritic joint and in the presence of body fluids acts like a battery and released gold ions continuously, thus relieving pain. However, the therapy is quite
expensive.
This invention envisages the use of gold nano particles as a safe substitute for suvarna bhasma having the same if not better properties and which can be made inexpensively very quickly and does not require the use of mercury, arsenic or sulfur in its manufacture.
According to this invention there is provided a formulation containing gold nano particles stabilized by an aqueous extract of biological tissue , said particles dispersed in a carrier in which the concentration of the gold nano particles ranges between 10 to 250 ppm.
The method of making the biologically stabilized gold nano particles has been described in detail in our copending patent application no: 550/MUM/2004 and involves a process for making gold sub micron scale particles comprising the steps of:
(a) dissolving a salt of the gold in water having conductivity less than 3 micro Siemens to obtain a solution in which the concentration of gold ions is in the range of 10, 000 to 25 000 ppm ,
(b) preparing a fresh filtered aqueous solution of biological tissue extract;
(c) diluting the aqueous solution with deionized water in the ratio ranging from 1:1 to 1: 25 to form a solution having a open circuit potential + 0.1 and +0.2 volt and a pH between 5.5 to 7.5 and a total organic carbon content of at least 7,500 ppm;

(d)maintaining the said aqueous solution under continuous agitation at a temperature between 20 and 30 degrees Celsius;
(e) inoculating a minute quantity of the gold salt solution in the said aqueous extract solution under continuous agitation such that the final concentration of the metal ion in the reaction mixture is in the range of ] 0 to 250 ppm;
(f) continuing the agitation for a period of 30 minutes to 5 hours to obtain a colloidal suspension of gold nano particles;
(g)separating the nano particles from colloidal suspension by a known process such as centrifugation.
The carrier may be a deionized water solution for injection or drops, an aerosol spray for inhalation, a granule, a globule or tableting formulation
Typically the biological tissue is derived from macerated plant cells selected from a group of plants consisting of: Alfalfa (Medicago sativa), Blood wort (Achillea millefolium), Garlic (Allium sativum) Indian aloe(Aloe barbadenesis), Celery(Apium graveolens) Ginger(Zingiber officinale), Castor seeds(Ricinus communis) Chirayata(Swertia chirayata), Colchium(Colchicum luteum) Gokulakanta(Hygrophila spinosa), Indian Gooseberry(Emblica officinalis), Indian Sarsaparilla (Hemidesmus indicus), Indian Senna(Cassia augustifolia), Indian Squill (Urginea indica), Lead wort(Plumbago zeylanica), Lemon Grass (Cymbopogon citratus), Madhuca(Madhuca indica), Nutmeg(Myristica fragrans), Pepper(Piper nigrum), Rosemary(Rosamarinus officinalis), Saffron(Crocus sativus) Sage(Salvia officinalis), Turpeth(Operculina turpethum), Vasaka(Adhatoda vasica), Winter cherry(Withania somnifera) Pergularia(Pergularia extensa), Onion(Allium cepa), Coriander(Coriandrum sativum,Linn), Turmeric(Curcuma longa), and

Tinospora cordifolia, Jatamansi [nordosstachys jatamansi], shatavari [asparagus racemosus] , triphala, brhami[ bocopa monnieri], Kappikachhu [Mucuna pruriens], deadly nighshade [atropa belladonna], and Centella asiatica.
The invention will now be described with reference to the accompanying text and drawings.
During the course of this invention ten gold bhasma samples available commercially very chemically analyzed and were found to have average gold concentration in the region of 82%. In addition elements such as iron (2%), sulfur (7%), oxygen (8%), arsenic (0.7%) and mercury (0.3%) were detected. The sizes of gold particles in the bhasma were analyzed by scanning electron microscopy and atomic force microscopy. Figure 1 in the accompanying drawings is a three dimensional atomic force microscope image of gold particles seen in one of the commercially obtained gold bhasma and figure 2 in the accompanying drawings is a gross graphical representation of particle size found in another gold bhasma sample. Figure 3 in the accompanying drawings is a three dimensional AFM image of yet another commercially obtained sample of gold bhasma showing gold particles.
It was seen that the sizes of gold particles in the commercially obtained suvarna bhasma ranged between 23 nm and almost 250nm with the majority being above 150 nm. These data indicate that the well documented therapeutic activity exhibited by suvarna bhasma is chiefly because of its gold content in the form sub micron size particles. It was therefore envisaged that if the gold particles in suvarna bhasma are replaced by gold nano particles biologically stabilized and therefore

biologically compatible, the resulting formulation will exhibit similar therapeutic activity.
To test this hypothesis during the course of present invention a comparative study on in vitro protease inhibition activity of suvarna bhasma and gold nano particles was carried out. Protease inhibition activity was selected because a variety of proteases such as matrix metallo proteinases [MMP], collagenases are overtly implicated in rheumatoid diseases. Therefore, protease inhibition is considered the most important parameter of therapeutic activity during the study.
Figure 4 in the accompanying drawings is a table showing the comparative percentage inhibition activity of gold nano particles in accordance with this invention, gold nano particles made conventionally by a chemical capping process and the activity of gold in gold bhasma.
As can be seen in the table in figure 4 of the drawings, the protease inhibition activity of the gold nano particles in accordance with this invention is favourably comparable with the activity of gold bhasma and in some cases even significantly better than gold bhasma.
A careful selection from the group of plant tissue from the list of plant described above is made and the gold nano particles can be used for specific disorders. For instance, a combination of aqueous extracts of withania somnifera, ginger, turmeric, pepper and tinospora cordifolia may be used in combination as the stabilizing solution for making a drug for the treatment of arthritis. Similarly, a combination of withania somnifera, brahmi, triphala, and centella asiatica may give better results in treating neuro pathological disorders.

Thus the gold nano particles biologically stabilized is a safefnon arsenic containing] substitute for gold bhasma with comparable activity.

We Claim:

[ 1] An arsenic free formulation for suvarna bhasma comprising
(.1) biologically stabilized gold nano particles in the size range of 1 to
100 mm; and
(2) a carrier iin which the concentration of the said biologically
stabllized gold nano particles is in the range of 80%.
[2] An arsenic free formulation as claimed in claim 1, in which the gold nano particles are stabilized biologically with an aqueous solution of macerated plant tissue cells .
[3] An arsenic free formulation as claimed in claim 2, in which the aqueous solution is diluted in deionised water in the up to ten folds.
[4] An arsenic free formulation as claimed in claim 2, in which the plant tissue is at least one plant tissue selected from a group of plant tissues which include leaves, roots, stems, flowers and fruits of the following plants Alfalfa (Medicagosativa), Blood wort (Achillea millefolium), Garlic (Allium sativum) Indian aloe(Aloe barbadenesis), Celery(Apium _ Igraveolens) Ginger(Zingiber officinale), Castor seeds(Ricinus communis) ' Chirayata(Swertia_chirayata)3 Colchium(CoIchicum luteum) Gokulakanta(Hygrophila spinosa), Indian Gooseberry(Emblica officinalis), Indian Sarsaparilla(Hemidesinus Jndicus), Indian Senna(Cassia augustifolia), Indian Squill(Urginea indica), Lead wort(Plumbago zeylanica), Lemon Grass(Cymbopogon citrarus), Madhuca(Madhuca indica), Nutmeg(Myristica fragrans), Pepper(Piper nigrum), Rosemary(Rosamarinus officinalis), Saffron(Crocus sativus)

Sage(Salvia officinalis), Turpeth(Operculina turpethum),
Vasaka(Adhatoda vasica), Winter cherry(Withania somnifera) Pergularia (Pergularia extensa), Onion(Allium cepa), Coriander(Coriandrum
satiyum,Linn), Turmeric(Curcuma longa), and Tinospora cordifolia, Jatamansi [nordosstachys jatamansi"], shatavari La'sparagus racemosus] , triphala, brhami[bocopa monnieri], Kappikachhu [Mucuna pruriens], deadly nighshade [atropa belladonna], and Centella asiatica.
[5] An arsenic free formulation as claimed in claim 1 or 2, in which the carrier is a liqujd, suspension, aerosol spray granules, globules, dry powder, and a tableting formulation .
[6] A method of making an antimicrobial formulation.aecording to any one of the preceding claims, which includes the steps of (1) making a carrier selected from a group of carriers which includes a liquid, suspension, aerosol spray granules, globules, dry powder,
and a tableting formulation in a conventional manner,
(2) making an aqueous dispersion of biologically stabilized gold nano
particles in the size range of 1 to 100 nm;
(3) mixing a dispensed quantity of the said aqueous dispersion in the
said carrier to form a homogenous matrix in which the
concentration of the biologically stabilized gold nano particles is
in the range of 10,000 to 25, 000 ppm.
[7] A method of making an arsenic free gold formulation as claimed in claim 6, in which the aqueous dispersion.of biologically stabilized gold nano particles is made by the steps of

(a) dissolving a salt of gold in water having conductivity less than 3
micro Siemens to obtain a solution in which the concentration of
gold ions is in the range of 50 000 ppm;
(b) preparing a fresh filtered aqueous solution of biological tissue
extract;
(c) diluting the aqueous solution with deionized water upto ten fold to
form a solution having an open circuit potential between + 0.2 and
+0.2 volt and a pH between 5.5 to 7.5 and total organic carbon
content at least 7,500 ppm;
(d) maintaining the said aqueous solution under continuous agitation at
a temperature between 20 and 30 degrees Celsius;
(e) inoculating gold salt solution in the said aqueous extract solution
under continuous agitation such that the final concentration of the
metal ion in the reaction mixture is in the range of 10 000 to 25
000 ppm;
(f) continuing the agitation for a period of 30 minutes to 3 hours ; to;
obtain a colloidal suspension of gold nano particles;
(g)separating the nano particles from colloidal suspension by a known process such as centrifugation.

Documents:

http://ipindiaonline.gov.in/patentsearch/GrantedSearch/viewdoc.aspx?id=FleQJ37jOoVxiZrMb4EL9A==&loc=vsnutRQWHdTHa1EUofPtPQ==

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Patent Number

269768

Indian Patent Application Number

551/MUM/2004

PG Journal Number

45/2015

Publication Date

06-Nov-2015

Grant Date

05-Nov-2015

Date of Filing

12-May-2004

Name of Patentee

NANO CUTTING EDGE TECHOLOGY PVT. LTD.

Applicant Address

D. LAD PATH, KALACHOWKI, MUMBAI 400033

Inventors:

#	Inventor's Name	Inventor's Address
1	KISHORE MADHUKAR PAKNIKAR	AGHARKAR RESEARCH INSTITUTE OF MAHARASHTRA ASSOCIATION FOR THE CULTIVATION OF SCIENCE G.G.AGARKAR ROAD, PUNE-411004.

PCT International Classification Number

B01J2/00 B22F9/24

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA