Anti AIDS Drug

AIDS Protocol Version 1.3 Confidential

Effect of An Ayurvedic Herbomineral preparation by Divya

Pharmacy in the treatment of AIDS (Ojakshaya): A
randomized controlled comparative study

Avnish K. Upadhyay

Department of Clinical Research

Divya Yog Mandir Trust, Patanjali Yog Peeth , Haridwar

September 2007

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TABLE OF CONTENTS

1. INTRODUCTION

1.1. Background
1.2. Hypothesis

2. STUDY OBJECTIVES

3. STUDY DESIGN

3.1. Study population

3.1.1. Inclusion Criteria
3.1.2. Exclusion Criteria
3.2. Study Observations
3.2.1. Screening Visit
3.2.2. Visit One
3.2.3. Subsequent two monthly visits

4. PATIENT WITHDRAWAL

5. TREATMENT ADMINISTERED

5.1. Randomization of Subjects

5.2. Dosage and Administration
5.2.1. Control/Conventional Regime
5.2.2. Ayurvedic Preparation

6. EFFICACY VARIABLES

6.1. Primary Endpoints

6.2. Secondary Endpoints

7. DATA ANALYSIS METHODS

7.1. Sample Size

7.2. Randomization
7.3. General Consideration
7.4. Statistical Methods

8. DATA COLLECTION

8.1 Demographics

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9. CLINICAL AND LAB PROCEDURES

9.1.
9.2.
9.3. Clinical Laboratory Assesments

10. REFERENCES

11. TABLE. TIME AND EVENT SCHEDULE

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1. INTRODUCTION

1.1. Background

Acquired immune deficiency syndrome or acquired immunodeficiency syndrome (AIDS

or Aids) is a collection of symptoms and infections resulting from the specific damage to
the immune system caused by the human immunodeficiency virus (HIV) in humans,[1]
and similar viruses in other species (SIV, FIV, etc.). The late stage of the condition leaves
individuals prone to opportunistic infections and tumors. Although treatments for AIDS
and HIV exist to slow the virus' progression, there is no known cure. HIV, et al., are
transmitted through direct contact of a mucous membrane or the bloodstream with a
bodily fluid containing HIV, such as blood, semen, vaginal fluid, preseminal fluid, and
breast milk.[2][3] This transmission can come in the form of anal, vaginal or oral sex, blood
transfusion, contaminated hypodermic needles, exchange between mother and baby
during pregnancy, childbirth, or breastfeeding, or other exposure to one of the above
bodily fluids.

Most researchers believe that HIV originated in sub-Saharan Africa during the twentieth
century;[4] it is now a pandemic, with an estimated 38.6 million people now living with
the disease worldwide.[5] As of January 2006, the Joint United Nations Programme on
HIV/AIDS (UNAIDS) and the World Health Organization (WHO) estimate that AIDS
has killed more than 25 million people since it was first recognized on June 5, 1981,
making it one of the most destructive epidemics in recorded history. In 2005 alone, AIDS
claimed an estimated 2.4–3.3 million lives, of which more than 570,000 were children. [5]
A third of these deaths are occurring in sub-Saharan Africa, retarding economic growth
and destroying human capital. Antiretroviral treatment reduces both the mortality and the
morbidity of HIV infection, but routine access to antiretroviral medication is not
available in all countries.[6] HIV/AIDS stigma is more severe than that associated with
other life-threatening conditions and extends beyond the disease itself to providers and
even volunteers involved with the care of people living with HIV.

AIDS is the most severe acceleration of infection with HIV. HIV is a retrovirus that
primarily infects vital organs of the human immune system such as CD4+ T cells (a
subset of T cells), macrophages and dendritic cells. It directly and indirectly destroys
CD4+ T cells. CD4+ T cells are required for the proper functioning of the immune system.
When HIV kills CD4+ T cells so that there are fewer than 200 CD4+ T cells per microliter
(µL) of blood, cellular immunity is lost, leading to the condition known as AIDS. Acute
HIV infection progresses over time to clinical latent HIV infection and then to early
symptomatic HIV infection and later to AIDS, which is identified on the basis of the
amount of CD4+ T cells in the blood and the presence of certain infections.

In the absence of antiretroviral therapy, the median time of progression from HIV
infection to AIDS is nine to ten years, and the median survival time after developing
AIDS is only 9.2 months.[7] However, the rate of clinical disease progression varies
widely between individuals, from two weeks up to 20 years. Many factors affect the rate

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of progression. These include factors that influence the body's ability to defend against
HIV such as the infected person's general immune function.[8][9] Older people have weaker
immune systems, and therefore have a greater risk of rapid disease progression than
younger people. Poor access to health care and the existence of coexisting infections such
as tuberculosis also may predispose people to faster disease progression.[7][10][11] The
infected person's genetic inheritance plays an important role and some people are
resistant to certain strains of HIV. An example of this is people with the CCR5-Δ32
mutation are resistant to infection with certain strains of HIV.[12] HIV is genetically
variable and exists as different strains, which cause different rates of clinical disease
progression.[13][14][15] The use of highly active antiretroviral therapy prolongs both the
median time of progression to AIDS and the median survival time.

According to Ayurveda, AIDS is primarily a disease of low ojas (immunity), the vital sap
of the body. Ojas provides the physical and mental strength to resist disease. With a
lowered immune system the body is susceptible to a host of opportune infections and
disorders. There are millions of bacteria and viruses in our environment, many of which
are contagious, that the body successfully fights off everyday. But in a condition of low
ojas or immunity level, these organisms are allowed to remain in the body, reproduce and
cause disease. Most of the symptoms associated with AIDS are similar to a disease
known in Ayurveda as ojaskshaya and detailed etiology and treatment methods are
available.

Ineffective metabolism of food produces toxins that pollute the system and reduce body
immunity (ojas). Stress, worry, overwork and excessive sexual indulgence also decrease
ojas. If this diminished state of vitality is allowed to continue, coupled with high-risk
exposure to infection, the body stands no chance of resisting disease. When all seven
dhatus or body tissues are deteriorated and ojas is reduced-disorders symptomatic of
AIDS can occur.

Ayurveda has many highly effective therapies for AIDS. There is a branch of Ayurveda
known as rasayana, which specifically deals with increasing immunity and vitality
through the use of various herbs, minerals and Ayurvedic techniques and practices. The
line of treatment aims at the relief of current symptoms while addressing the underlying
cause. After initial detoxification, the strength, immunity and vitality of the patient are
increased through rasayanic therapies, which after a period of time can bring about a
complete cure.

The Symptoms and causative factors and treatment for the latter are found mentioned in
many ancient Ayurvedic tests like, Charaka Samhitha, Bhavaprakasha,Vaidyachintamani
and Chakradatta etc., Shosha is another condition, which results from loss of energy that
is similar to AIDS.

The disease is caused by the dominant kapha dosha along with the other doshas tends to
block the path for the flow of rasadi dhatus in their respective locations thus resulting in
the deterioration of saptha dhatu Rasa(plasma), Rakta (blood cells),Mansa ( muscular

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tissue), Meda (adipose tissue), Asthi (bony tissue), Majja (bone marrow) and the Shukra
(reproductive tissue). thus resulting into the disease. (16)

The major symptoms are:

• loss of appetite, drastic loss of weight-emaciation

• fatigue and lethargy
• susceptibility to allergies and contagious diseases
• skin irritations
• bronchial disorders, often leading to tuberculosis of the lungs
• damage to intestinal flora resulting in diarrhoea, dysentery, gastritis and
• wide fluctuations in body temperature. prolonged fever
• sleeplessness etc. The symptoms of AIDS are primarily the result of conditions
that do not normally develop in individuals with healthy immune systems. Most
of these conditions are infections caused by bacteria, viruses, fungi and parasites
that are normally controlled by the elements of the immune system that HIV
damages. Opportunistic infections are common in people with AIDS. (17)

Scientific Review:

Administration of an extract from the powdered root of the plant Withania somnifera was
found to stimulate immunological activity in Babl/c mice. Treatment with five doses of
Withania root extract (20 mg/dose/animal; i.p.) was found to enhance the total WBC
count (17 125 cells/mm3) on 10th day. Bone marrow cellularity (27×106 cells/femur) as
well as α-esterase positive cell number (1800/4000 cells) also increased significantly
(P<0.001) after the administration of Withania extract. Treatment with Withania extract
along with the antigen (SRBC) produced an enhancement in the circulating antibody titre
and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC
(985 PFC/106 spleen cells) was obtained on the fourth day. Withania extract inhibited
delayed type hypersentivity reaction in mice (Mantoux test). Administration of Withania
extract also showed an enhancement in phagocytic activity of peritoneal macrophages
(76.5 pigmented cells/200) when compared to control (31.5/200 cells) in mice. These
results confirm the immunomodulatory activity of W. somnifera extract, which is a
known immunomodulator in indigenous medicine.(18)

Withania somnifera is an Indian medicinal plant used widely in the treatment of many
clinical conditions in India. Its antistressor properties have been investigated in this study
using adult Wistar strain albino rats and cold water swimming stress test. The results
indicate that the drug treated animals show better stress tolerance.(19)

The antioxidant activity of Withania somnifera (WS) glycowithanolides was assessed in

chronic footshock stress induced changes in rat brain frontal cortex and striatum. The
stress procedure, given once daily for 21 days, induced an increase in superoxide
dismutase (SOD) and lipid peroxidation (LPO) activity, with concomitant decrease in
catalase (CAT) and glutathione peroxidase (GPX) activities in both the brain regions. WS
glycowithanolides (WSG), administered orally 1 h prior to the stress procedure for 21

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days, in the doses of 10, 20 and 50 mg/kg, induced a dose-related reversal of the stress
effects. Thus, WSG tended to normalise the augmented SOD and LPO activities and
enhanced the activities of CAT and GPX. The results indicate that, at least part of chronic
stress-induced pathology may be due to oxidative stress, which is mitigated by WSG,
lending support to the clinical use of the plant as an antistress adaptogen.(20)

Antigenotoxic properties and the possible mechanisms of water extracts from Cassia tora
L. (WECT) treated with different degrees of roasting (unroasted and roasted at 150 and
250 degrees C) were evaluated by the Ames Salmonella/microsome test and the Comet
assay. Results indicated that WECT, especially unroasted C. tora (WEUCT), markedly
suppressed the mutagenicity of 2-amino-6- methyldipyrido(1,2-a:3':2'-d) imidazole (Glu-
P-1) and 3-amino-1, 4-dimethyl-5H-pyrido (4,3-b)indole (Trp-P-1). In the Comet assay
performed on human lymphocytes, WECT exhibited significant protective effect on Trp-
P-1-mediated DNA damage followed the order of unroasted (55%) > roasted at 150
degrees C (42% ) > roasted at 250 degrees C (29%). Pre-treatment of the lymphocytes
with WEUCT resulted in 30% repression of DNA damage. However, no significant effect
on excision-repair system was found during DNA damage expression time in post-
treatment scheme (p>0.05). WEUCT showed 84% scavenging effect on oxygen free
radicals generated in the activation process of mutagen detected by electron paramagentic
resonance system. Two possible mechanisms were considered: (1) neutralization the
reactive intermediate of Trp-P-1; and (2) protecting cells directly as an antioxidant that
scavenge the oxygen radicals from the activation process of mutagen. The individual
anthraquinone content in extracts of C. tora was measured by HPLC. Three
anthraquinones, chrysophanol, emodin and rhein, have been detected under experimental
conditions. The anthraquinone content decreased with increased roasting temperature.
Each of these anthraquinones demonstrated significant antigenotoxicity against Trp-P-1
in the Comet assay. In conclusion, our data suggest that the decrease in antigenotoxic
potency of roasted C. tora was related to the reduction in their anthraquinones.(21)

The effect of water extracts from Cassia tora L. (WECT) were tested for their effects on
DNA damage in human hepatoma cell line HepG2. DNA damage in HepG2 cells could
be reduced by WECT in a dose-dependent manner.(22)

The antimutagenic activity of a methanol extract of Cassia tora seeds against aflatoxin
B1(AFB1) was demonstrated. Column chromatography using silica gel yielded pure
chrysophanol, chryso-obtusin, and aurantio-obtusin from the CH2Cl2 fraction and
cassiaside and rubro-fusarin gentiobioside from the n-BuOH fraction. Each of these
compounds demonstrated significant antimutagenic activity.(23)

The fruits extracts of Emblica officinalis (Amla) has been reported to have strong anti-
oxidant properties. There is a paucity of studies on the immunomodulatory properties of
fruit extracts of Amla in immuno-compromised states, with the emphasis on
lymphocytes. Therefore, the aim of the study was to determine the anti-oxidant and
immunomodulatory properties of Amla using chromium (VI) as an immunosuppressive
agent. Chromium (Cr) treatment results in enhanced cytotoxicity, free radical production,
lipid peroxidation and decreased glutathione peroxidase (GPx) activity and diminished

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glutathione (GSH) levels. There was a significant inhibition of both lipopolysaccharide

and concanavalin-A-stimulated lymphocyte proliferation. Chromium also inhibited Con
A stimulated interleukin-2 and γ-interferon production significantly. Further, there was
enhanced apoptosis and DNA fragmentation in the presence of Cr. Amla significantly
inhibited Cr-induced free radical production and restored the anti-oxidant status back to
control level. Amla also inhibited apoptosis and DNA fragmentation induced by Cr.
Interestingly, Amla relieved the immunosuppressive effects of Cr on lymphocyte
proliferation and even restored the IL-2 and γ-IFN production considerably.(24)

Cyclophosphamide (CP) is one of the most popular alkylating anticancer drugs in spite of
its toxic side effects including immunotoxicity, hematotoxicity, mutagenicity and a host
of others. The present study was undertaken to assess the protective effects of total
aqueous extract of a medicinal plant, Indian gooseberry (Emblica officinalis Gaertn.) in
mice treated with CP. These protective effects were studied on immunological parameters
and kidney and liver antioxidants. Plant extract treatment at a dose of 100 mg/kg body
weight per os (p.o.) for 10 days resulted in the modulation of these parameters in normal
as well as CP (50 mg/kg)-treated animals. Plant extract in particular was very effective in
reducing CP-induced suppression of humoral immunity. Plant extract treatment in normal
animals modulated certain antioxidants of kidney and liver. In CP-exposed animals, plant
pretreatment provided protection to antioxidants of kidney. Not only were the reduced
glutathione levels significantly (p<0.001) increased but plant extract treatment resulted in
restoration of antioxidant enzymes in CP-treated animals. It is suggested that E.
officinalis or its medicinal preparations may prove to be useful as a component of
combination therapy in cancer patients under CP treatment regimen.(25)

Diabetic patients with foot ulcers on T. cordifolia as an adjuvant therapy showed

significantly better final outcome with improvement in wound healing. Reduced
debridements and improved phagocytosis were statistically significant, indicating
beneficial effects of immunomodulation for ulcer healing.(26)

Effect of Tinospora cordifolia extract on modulation of hepatoprotective and

immunostimulatory functions in carbon tetrachloride (CCl4) intoxicated mature rats is
reported here. Administration of CCl4 (0.7 ml/kg body weight for 7 days) produces
damage in the liver as evident by estimation of enzymes such as serum glutamate
oxaloacetate transaminase (SGOT), serum glutamate pyruvate transminase (SGPT) and
alkaline phosphatase (ALP) as well as serum bilirubin level. CCl4 administration also
causes immunosuppressive effects as indicated by phagocytic capacity, chemotactic
migration and cell adhesiveness of rat peritoneal macrophages. However, treatment with
T. cordifolia extract (100 mg/kg body weight for 15 days) in CCl4 intoxicated rats was
found to protect the liver, as indicated by enzyme level in serum. A significant reduction
in serum levels of SGOT, SGPT, ALP, bilirubin were observed following T. cordifolia
treatment during CCl4 intoxication. Treatment with T. cordifolia extract also deleted the
immunosuppressive effect of CCl4, since a significant increment in the functional
capacities of rat peritoneal macrophages (PM.PHI.) was observed following T. cordifolia
treatment. The results of our experiment suggest that treatment by T. cordifolia extract

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may be the critical remedy for the adverse effect of CCl4 in liver function as well as
immune functions.(27)

The immunomodulatory properties of NIM-76 have been described in this paper. Pre-
treatment of rats with a single i.p. injection of NIM-76 resulted in an increase in
polymorphonuclear (PMN) leukocytes with a concomitant decrease in lymphocyte
counts. The immunomodulatory activity of NIM-76 was found to be concentration-
dependent. At 120 mg/kg body weight, there was an enhanced macrophage activity and
lymphocyte proliferation response, while the humoral component of immunity was
unaffected. At higher concentrations of NIM-76 (300 mg/kg body weight), there was a
stimulation of mitogen-induced lymphocyte proliferation, while macrophage activity
remained unaffected. However, a fall in primary and secondary antibody titres was
observed. The study indicates that NIM-76 acts through cell-mediated mechanisms by
activating macrophages and lymphocytes.(28)

The alkaloidal fraction of Boerhaavia diffusa was studied for its effect on cellular and
humoral functions in mice. Oral administration of the fraction (25–100 mg/kg)
significantly inhibited SRBC-induced delayed hypersensitivity reactions in mice.
However, the inhibition was observed only during post-immunisation drug treatment,
while no effect during pre-immunisation drug treatment was observed. A significant
dose-related increase in antibody titre was observed during pre- and post-immunisation
treatment. The alkaloidal fraction failed to show any blastogenic responsiveness of
murine splenocytes to Concanvalin A (Con A) and lipopolysaccharide (LPS). Similarly,
it did not display any mitogenic activity. Thus, the present study has shown the in vivo
immunostimulatory activity of B. diffusa alkaloidal fraction without an in vitro effect.(29)

Celastrus paniculatus L. (Celastraceae) (CP), Picrorhiza kurroa L. (Scrophulariaceae)

(PK) and Withania somnifera L. (Solanaceae) (WS) are Indian medicinal plants having a
remarkable reputation, as a factor of health care, among the indigenous medical
practitioners. The plants exhibit varying degrees of therapeutic value some of which
useful in the treatment of cognitive dysfunction, epilepsy, insomnia, rheumatism, gout,
dyspepsia.(30)

Psoralea corylifolia seed extract has been found to stimulate the immune system in mice.
Administration of the extract was found to inhibit EAC ascitic tumour growth and
stimulate natural killer cell activity, antibody-dependent cellular cytotoxicity, antibody-
forming cells and the antibody complement-mediated cytotoxicity during tumour
development.(31)

Methanolic extracts of 20 medicinal plants were screened at 1-10 mg/ml for in vitro
macrofilaricidal activity by worm motility assay against adult Setaria digitata, the cattle
filarial worm. Four plant extracts showed macrofilaricidal activity by worm motility at
concentrations below 4 mg/ml and an incubation period of 100 min. Complete inhibition
of worm motility and subsequent mortality was observed at 3, 2, 1 and 1 mg/ml,
respectively, for Centratherum anthelminticum, Cedrus deodara, Sphaeranthus indicus

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and Ricinus communis. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide

(MTT) reduction assay was carried out at 1 mg ml−1 and 4-h incubation period, and the
results showed that C. deodara, R. communis, S. indicus and C. anthelminticum exhibited
86.56, 72.39, 61.20 and 43.15% inhibition respectively in formazan formation compared
to the control. (32)

Curcuma longa (turmeric) is a well-known indigenous herbal medicine. The aqueous

extracts, when administered orally to the mice from 140 to 560 mg/kg for 14 days, were
able to elicit dose-dependent relation of immobility reduction in the tail suspension test
and the forced swimming test in mice. The effects of the extracts at the dose of 560
mg/kg were more potent than that of reference antidepressant fluoxetine. The extracts, at
the dose of 140 mg/kg or above for 14 days, significantly inhibited the monoamine
oxidize A (MAO) activity in mouse whole brain at a dose-dependent manner, however,
oral administration of the extract only at a dose of 560 mg/kg produced observable MAO
B inhibitory activity in animal brain. Fluoxetine showed only a tendency to inhibit MAO
A and B activity in animal brain in the study. Neither the extracts of C. longa nor
fluoxetine, at the doses tested, produced significant effects on locomotor activity. These
results demonstrated that C. longa had specifically antidepressant effects in vivo. The
activity of C. longa in antidepression may mediated in part through MAO A inhibition in
mouse brain.(33)

Curcumin (diferuloylmethane), found in the spice turmeric, exhibits anti-inflammatory,

antioxidant, and chemopreventive activities. However, the effect of curcumin on the
immunological responses largely remains unknown. In this study we have investigated
the effect of curcumin on mitogen (phytohaemagglutinin; PHA) stimulated T-cell
proliferation, natural killer (NK) cell cytotoxicity, production of cytokines by human
peripheral blood mononuclear cells (PBMCs), nitric oxide (NO) production in mouse
macrophage cells, RAW-264.7. Furthermore, we have carried out an electromobility shift
assay to elucidate the mechanism of action of curcumin at DNA protein interaction level.
We observed that curcumin inhibits PHA-induced T-cell proliferation, interleukin-2
production, NO generation, and lipopolysachharide-induced nuclear factor- B (NF- B)
and augments NK cell cytotoxicity. Our results suggest that curcumin most likely inhibits
cell proliferation and cytokine production by inhibiting NF- B target genes involved in
the induction of these immune parameters.(34)

Sida cordifolia L. (Malvaceae) is used in folk medicine for the treatment of inflammation
of the oral mucosa, blenorrhea, asthmatic bronchitis and nasal congestion. The anti-
inflammatory, analgesic effects and acute toxicity of an aqueous extract of S. cordifolia
were evaluated in animal models. The extract was prepared using leaves collected before
the flowering period. The aqueous extract (AE) showed a significant inhibition of
carrageenin-induced rat paw edema at a dose of 400 mg/kg administered orally, but did
not block the edema induced by arachidonic acid. The AE also increased the latency
period for mice in the hot plate test, and inhibited the number of writhes produced by
acetic acid at the oral dose of 400 mg/kg. The aqueous extract of S. cordifolia showed
low acute toxicity in mice.(35)

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Fourteen participants (25%) reported use of one or more herbs (range 1-8) in the prior
three months, at a mean cost of $49 per month (range 0-150). 28% were unable to
identify the herb that they used. Use of herbs was not correlated with a diagnosis of AIDS
or ARC, nor with the number of symptoms reported. Of those patients taking herbs 57%
reported that they obtained their information from alternative healers such as herbal
practitioners, acupuncturists, chiropractors, or homeopaths, and 36% actually received
their herbs from an alternative healer. Of those taking herbs, 21% were involved in
clinical drug trials. Several patients reported taking herbs in amounts at which
pharmacologic or even toxic effects have been seen in case reports. Potential adverse
effects of these herbal medications include anaphylactic and anaphylactoid reactions, skin
rashes, GI irritation and diarrhea, thrombocytopenia and coagulopathies, changes in
mental status, hepatotoxicity, and electrolyte disturbances. These can also be symptoms
of HIV disorders or side effects of drugs given to treat HIV infected patients.: The
prevalence of medicinal herb use underscores the need for physicians treating AIDS
patients, and those running clinical trials to obtain specific information on patients' use of
herbs. Patient care and clinical trials could be confounded by the use of herbs whose
pharmacologic effects can resemble commonly occurring symptoms in HIV disorders or
side effects of prescribed or protocol medications(36)

Herbs have been used as food and for medicinal purposes for centuries. Research interest
has focused on various herbs that possess hypolipidemic, antiplatelet, antitumor, or
immune-stimulating properties that may be useful adjuncts in helping reduce the risk of
cardiovascular disease and cancer. In different herbs, a wide variety of active
phytochemicals, including the flavonoids, terpenoids, lignans, sulfides, polyphenolics,
carotenoids, coumarins, saponins, plant sterols, curcumins, and phthalides have been
identified. Several of these phytochemicals either inhibit nitrosation or the formation of
DNA adducts or stimulate the activity of protective enzymes such as the Phase II enzyme
glutathione transferase (EC 2.5.1.18). Research has centered around the biochemical
activity of the Allium sp. and the Labiatae, Umbelliferae, and Zingiberaceae families, as
well as flaxseed, licorice root, and green tea. Many of these herbs contain potent
antioxidant compounds that provide significant protection against chronic diseases. These
compounds may protect LDL cholesterol from oxidation, inhibit cyclooxygenase and
lipoxygenase enzymes, inhibit lipid peroxidation, or have antiviral or antitumor activity.
The volatile essential oils of commonly used culinary herbs, spices, and herbal teas inhibit
mevalonate synthesis and thereby suppress cholesterol synthesis and tumor growth.(37)

Various ingredients used in present herbomineral preparation are found very much
effective in immune related disorders. (38-46)

1.2. Hypothesis

Many ayurvedic formulations given in traditional texts used by ayurvedic practitioners

for long times like Heerak Bhasma, Ras Manikya, Swarn Basant Malti, Amrita sat etc.
are looking very much useful in different types of immune related disorders, may be due
to lekhan properties of these drugs.

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2. STUDY OBJECTIVES

The assessment of relative importance of Effect of An Ayurvedic Herbomineral

preparation by Divya Pharmacy in the treatment of AIDS (Ojakshaya). The result of this
study will decide long term study on AIDS patients.

3. STUDY DESIGN

******* newly diagnosed AIDS patients will be recruited and randomized in a 1 Control
Group : 1 Ayurvedic regime
fashion and followed for at least two years.

The following pathological parameters will be investigated after 45 days, 90 days and
every 3 months.

- Hb %

- WBC (White blood cells) count

- Platelets counts

Other Investigations:

- ***********

- ***********

Treatment failure:

- Negative changes in parameters

- If patients condition not improving in sense of QoL etc.

Outcomes:

- Mortality

- Treatment failure

3.1. Study population

3.1.1. Inclusion Criteria

A subject will be included in the study only if all the following criteria apply:

Clinical diagnosis of HIV/AIDS

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Must be able to swallow tablets

Must be able to eat nutritional foods

Have HIV infection

Have signed consent of parent or guardian for patients under 21 years of age

Are able and willing to use the study drugs

Are to be followed at a participating clinical site

Children of any age (greater than 8 years old)

Patient who have given written informed consent.

3.1.2. Exclusion Criteria

A subject will not be eligible for inclusion in the study if any of the following criteria
apply:

Medical side effects

Are pregnant or breast feeding

Have a history of significant cardiac abnormalities or dysfunction

Are using anti retroviral drugs. (HIV patients are not included in the first batch.)

Are receiving certain drugs or treatments

Unable to be followed at a participating clinical center

Children less than 8 years old

Have any serious conditions (severe chronic staged AIDS cases) at study entry that might
affect the results of the study

Allergy to any of the study drugs or their formulations

Tobacco using patients will be excluded from these trials.Alcohol using patients will be
excluded from these trials.

Drug addicted patients will be excluded from these trials.

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Patients with significant psychiatric disorder or mental disability.

3.2. Study Observations

3.2.1. Screening Visit

A maximum of 7-8 days elapse between screening and the start of treatment. Patients will
be randomized and assigned an identification number during the screening visit.

The following procedure will be performed:

1. Patient must sign informed consent form.

2. Physical examination.

3. Collection of blood for cytochemistry, biochemistry and haematology analysis.

4. Record of Vital signs

The following information will be recorded:

1. Demographics including – Date of birth, Gender and race.

2. Height, Weight

3. Primary disease.

3.2.2. Visit One

Visit one will take place around 45 days after treatment starts.

The following procedure will be performed:

1. Physical examination.

2. Collection of blood for cytochemistry, biochemistry and haematology analysis.

3. Record of Vital signs

The following information will be recorded:

1. Date treatment started

2. Weight

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3.2.3. Subsequent two monthly visits

Treatment period continues for twelve months. Patients will be with drawn from the
study if they have life threatening problem.

The following procedure will be performed:

1. Physical examination.

2. Collection of blood for cytochemistry, biochemistry and haematology analysis.

3. Record of Vital signs

The following information will be recorded:

1. Treatment regime

2. Weight

4. PATIENT WITHDRAWAL

Patients are withdrawn from the study for any of the following reasons:

1. Completion of study

2. Patient preference

3. Death

4. Physician discretion

5. TREATMENT ADMINISTERED

5.1. Randomization of Subjects

Patients will be randomized 1:1

1 Mixture of certain drugs of Ayurveda widely used for different treatments)

1 Control Group

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5.2. Dosage and Administration

5.2.1. Control Group

5.2.2. Ayurvedic Preparation

Each 500 mg Capsule or Tablet contains

Name Botanical/English Name Qty. (mg.)
Extract of >>>>>0.45mg Each
1 Panwar Cassia tora --
Daruhaldi Barberis aristata --
Karanj Pongamia pinnata --
Amla Emblica officinalis --
Giloy Tinospora cordifolia --
Kutki Picrirhiza kurroa --
Bakuchi Phal Psoralia corylifolia --
Bahera Terminalia chebula --
Kali Jeeri Centratherum anthelminticum --
Choti Kateli Solanum surattense --
Haldi Curcuma longa --
Khair Acacia catechu --
Nimb Azadirachta indica --
Manjeeth Rubia cordifolia --
Chirayita Swertia chirayita --
Dronpushpi Leucas cephalotes --
Madhuyasthi Glycyrhiza glabra --
Indrayanmool Cirullus colocynthis --
Ashwagandha Withania somnifera --
Usba Smilex arnata --
Punarnavamool Boerhavia diffusa --
Bhuiamla Phyllanthus urinaria --
Makoy Solanum nigrum --
Bala Sida cordifolia --
Nirgundi Vitex nirgundo --
Powders of
2 Heerak Bhasma >>>>>3.3 mg.
Ras Manikya >>>>33.3 mg.
Swarn Makshik >>>>27.5 mg.
Swarn Basant Malti >>>>16.5 mg.
Amrita Sat >>>>53.3 mg.
Mukta Pishti >>>>27.5 mg.
Praval Panchamrit >>>>27.5 mg.
3 Kaishore Guggulu >>>>200 mg.
Arogyavardhini Vati >>>>100 mg.

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2 tab. Or capsule will be given to patients with 50ml. fresh juices of wheat grass and
guduchi - Tinosporsa cordifolia (25 ml. each) twice in a day half n hour before breakfast
and dinner.

6. EFFICACY VARIABLES

6.1. Primary Endpoints

6.2. Secondary Endpoints

7. DATA ANALYSIS METHODS

7.1. Sample Size

A minimum of ** patients will be recruited to commence the study in order to detect the
minimum relevant clinical difference at a statistical power of 80 % and p=0.05.

7.2. Randomization

Patients will be randomized 1:1

1 Mixture of certain drugs of Ayurveda widely used for different treatments)

1 Control Group

7.3. General Consideration

As this is a randomized controlled trial, the primary analysis will be an intend-to-treat

(ITT) analysis whereby all comparisons will be made on the basis of the treatment group
to which patients are initially randomized.

A secondary as-treated analysis will also be performed based solely on those patients
deemed to be evaluable throughout the study. This as-treated analysis will directly access
the effectiveness of treatment regime with respect to the primary and secondary outcome
variables.

7.4. Statistical Methods

Student‘t’ test and repeated measure ANOVA.

8. DATA COLLECTION

8.1. Demographics

Demographic measure includes age, gender and race.

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9. CLINICAL AND LAB PROCEDURES

9.1. Clinical Laboratory Assesments

1. Haematology

5ml blood in to EDTA will be screened for

RBC count

Hb%

WBC count

TLC

DLC

MCV

MCH

MCHC

Platelet count

2. Biochemistry

10ml blood in to heparinised tubes to screen for:

Serum uric acid

Glucose

Albumin

Total protein

Total bilirubin

Urea

Creatinine

Total cholesterol

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HDL

LDL

Triglyceride

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