T-AYU-HM™ PremiumAnti-Sickling Medicine for Management of Sickle

Cell Anemia.

Sicklecare TAYUHM Limited.

Improving Quality of Life…

T-AYU-HM™ Premium

Composition: Each T-AYU-HMTM Premium

tablet (300 mg) contains:

1 Calyx of Mica 25mg

2 Calyx of Iron 12.5mg

3 Terminala chebula 25mg

4 Zingiber officinale 25mg

5 Asparagus racemosus 25mg

6 Punica granatum 12.5mg

7 Myristica fragrans 25 mg

8 Piper longum 37.5 mg

9 Tinospora cordifolia 37.5 mg

10 Leptadinia reticulate 37.5 mg

In-Vitro RBC Sickling (Emmel’s Test) Results

The morphology ofSickle Cells that wereincubated underanaerobic conditionsin the absence orpresence of variousconcentration of T-AYU-HM™ Premiumcompared to 10mMVanillin (4-Hydroxy-3-methoxybenzaldehyde) for 24 hour.

- Ve Control + Ve Control

10 mM Vanillin as standard

25 µg/ml

T-Ayu-HM™ Pre

50 µg/ml

T-Ayu-HM™ Pre

100 µg/ml

T-Ayu-HM™ Pre

In-Vitro RBC Sickling (Emmel’s Test) Results

- Ve Control + Ve Control 10 mM Vanillin as standard

25 µg/ml

T-Ayu-HM™ Pre

50 µg/ml

T-Ayu-HM™ Pre

100 µg/ml

T-Ayu-HM™ Pre

500 µg/ml

T-Ayu-HM™ Pre

In-Vitro RBC Sickling (Emmel’s Test) Results

5

*

*

**

*

0

10

20

30

40

50

60

70

80

90

100

control 10 mMvanillinstardard

25 µg/mlHMP

50 µg/mlHMP

100 µg/mlHMP

500 µg/mlHMP

% S

ickle

d C

ells

Percent of Sickled cells in presence of T-AYU-HM™ Premium

n=5, * p<0.05 as compared to control samples

Treatment Control (PBS)10 mM

Vanillin

25 µg/ml

T-Ayu-HMP

50 µg/ml

T-Ayu-HMP

100 µg/ml

T-Ayu-HMP

500 µg/ml

T-Ayu-HMP

% Sickle Cells

(Mean ±SD)

67.94

±4.47

24.07

±10.66

55.22

±4.66

44.28

±5.77

39.34

±5.90

27.41

±8.12

% Inhibition

of sickling

(Mean ± SD)

-

64.62

±15.41

18.32

±10.11

34.51

±10.21

41.83

±10.33

59.85

±11.36

In-Vitro RBC Sickling (Emmel’s Test) Results

Treatment Control (PBS)10 mM

Vanillin

25 µg/ml

T-Ayu-HMP

50 µg/ml

T-Ayu-HMP

100 µg/ml

T-Ayu-HMP

500 µg/ml

T-Ayu-HMP

% Sickle Cells

(Mean ±SD)

67.94

±4.47

24.07

±10.66

55.22

±4.66

44.28

±5.77

39.34

±5.90

27.41

±8.12

% Inhibition

of sickling

(Mean ± SD)

-

64.62

±15.41

18.32

±10.11

34.51

±10.21

41.83

±10.33

59.85

±11.36

Oxidative Hemolysis of Sickle Red Blood Cells

-10.00

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

0 15 30 45 60 75 90 105 120

Perc

ent

Haem

oly

sis

Time in Min

Control

50 ug/ml T-Ayu-HM

100 ug/ml T-ayu-HM

500 ug/ml T-ayu-HM

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

90.00

100.00

Control 50 ug/ml T-Ayu-HM

100 ug/ml T-ayu-HM

500 ug/ml T-ayu-HM

Meth

aem

oglo

bin

(%

)

t-BOOH induced Formation of Methaemoglobin at 12 Hr

* indicates significantly different means from the control groups (P<0.05) analyzed by one-way ANOVA followed by Dunnett's

comparison test

*

*

Membrane Stabilization Activity byOsmotic Fragility Test

0.00

20.00

40.00

60.00

80.00

100.00

120.00

0.00 0.20 0.40 0.60 0.80 1.00

Perc

enta

ge H

aem

oly

sis

Concentration of NaCl Solution

control (PBS)

50 µg/ml T-Ayu-HM

100 µg/ml T-Ayu-HM

500 µg/ml T-Ayu-HM

10 mM Vanillin

0.45

0.47

0.49

0.51

0.53

0.55

0.57

0.59

control(PBS)

10 mMVanillin

50 µg/ml T-Ayu-HM

100 µg/mlT-Ayu-HM

500 µg/mlT-Ayu-HM

Conc.

of

NaC

l

Concentration at Haemolysis 50%

Stasis Induced Venous Thrombosis in SD Rats

0

1

2

3

4

5

6

7

8

9

10

Vehical Control 30 mg/ kgAspirin

100 mg/kgT-AYU-HM

300 mg/kgT-AYU-HM

500 mg/kgT-AYU-HM

Weig

ht

of

Thro

bis

(m

g)

Stasis Induced Venous Thrombosis in SD Rats

n=5, * p<0.05 as compared to control samples using Student’s t test

** *

Conclusion

The ability of T-AYU-HM Premium to normalize the sickle erythrocytes may represent a rational explanation for the use in treating sickle cell anemia.

Sickled RBCs showed a lesser degree of t-BOOH-induced haemolysis in treatment group than control groups and were more resistant to t-BOOH induced hemolysis.

T-AYU-HM Premium causes stabilization of the erythrocytes membrane, reflected in the decrease in osmotic fragility values.

T-Ayu-HM premium significantly shows decrease in thrombus formation.

The present investigation provides evidence to support the claims of theherbomineral formulation for treatment and management of sickle cellanemia

Discussion

• Literature survey revealed that herbomineral formulationsinclude various plant extracts and minerals which can beuseful in the complications of sickle cell disease.

For instance,

• Zingiber officinale contains essential oil which possess anti-inflammatory activity (Thomson et al., 2002).

• Punica Granatum containing brevifolin carboxylic acid,brevifolin, 7,8-Dihydroxycoumarin 7-Ellagic acid are knownto posses anti-oxidant and analgesic activities (Singh et al.,2002, Nawwar et al., 1994, Hussein et al., 1997).

• Myristica Fragrans shows potent antiplatelet activity,Antioxidant activity (Duan et al., 2009)

Discussion

• Piper longum shows inhibitory activity on plateletaggregation induced by collagen, arachidonic acid, andplatelet-activating factor (Park et al., 2007),

• Terminalia Chebula shows antioxidant properties , containspolyphenolic compounds like casuarinin, chebulanin,chebulinic acid or 1,6-di-O-galloyl-β -D-glucose (Cheng etal., 2003).

• Asparagus racemosus shows cytoprotective activity(Dahanukar et al., 1983) as well as antioxidant activity(Kamat et al., 2000),

• Mica Ash useful in anemia, splenomegaly (Desai, 2008), agingand general debility,

• Iron Ash used in treatment of iron deficiency anaemia(Pandit et al., 1999).

Application

o T-Ayu-HM™ Premium is being used in India by proficient Practitioners for thetreatment of sickle cell anemia

o Dramatic improvements in the patients conditions were observed with respect toReduction or freedom from symptoms

Hand & Chest syndrome

Avascular necrosis of femur (AVNF)

Relief from pain

Splenomegaly, jaudice, pallor, backache, abdominal colic, loss of appetite, and headache.

Improvement in hematological parameters

Need for Blood transfusions is also considerably reduced.

Summary of overall relief

It improves quality of life of the patient It improves general conditions of the patient It drastically reduces the need for repeated blood transfusions. It normalizes renal and liver functions it eliminates the need for splenectomy it strengthens the cardiac functions it eliminates chronic fatigue; patients feel more energetic it prevents kidney damage due to the disease it prevents formation of gall stones it improves RBC function it consists of a balanced mixture of time-tested and safe herbominerals without

any side effects it is manufactured as per FDA standards Each ingredient is subjected to physical and chemical analysis Its formulation is subjected to Heavy Metal Microbial testing Its safety for human use is ascertained by LD50 values

Dosage

• 2 tablets daily (Morning and Evening) or as directed by physician for HbSS.

• In severe condition 2 tablets twice daily for 120 days for HbSS.

• During Crisis 2 tablets twice daily with lifesavers medications.

• 1 tablet daily (Morning and Evening) for HbAS.• 2 tablets daily for complications of HbAS.

Warning : pregnant women, nursing mothers and children under 5 years have to be

prescribed by the attending doctors.

Indications• INDICATIONS AND USAGE: T-AYU-HM ™ Premium tablets are indicated mainly for sickle cell anemia

.The product is also useful in other types of anemia

• CONTRA INDICATIONS: T-AYU-HM ™ Premium tablets are contraindicated during pregnancy and lactation .

• WARNING AGAINST MISUSE: T-AYU-HM ™ Premium is indicated for a specific condition called sickle cell anemia and should be used as per the dosage recommended by the manufacturer; should not be used for conditions not recommended by the manufacturer.

• PRECAUTIONS: During the use of T-AYU-HM ™ Premium tablets, patients have to be carefully observed and any untoward reactions to be reported immediately to the attending Physician. Patients have to be carefully and continuously monitored by the attending Physician during the T-AYU-HM ™ Premium therapy. In severe cases of Sickle cell anemia there could be additional need for measures, medications, blood transfusions and even hospitalizations-which should be promptly resorted to.

• DOSAGE AND ADMINISTRATION: 2 tablets of 300mg twice a day or as directed by the Physician.

• ADVERSE REACTIONS: No specific adverse reaction was reported in patients who have used T-AYU-HM ™ Premium

• DRUG ABUSE AND DRUG DEPENDENCE: T-AYU-HM ™ Premium Tablets contain time tested herbal and Ayurvedic ingredients. There is no propensity to cause drug abuse and drug dependence. There is no report of drug abuse or drug dependence.

• SYMTOMS OF OVERDOSAGE AND ANTIDOTE: None.

References

• P.T. Mpiana, D.S. Tshibangua, O.M. Shetondea, K.N. Ngboluab “In vitro antidrepanocytary actvity(anti-sickle cell anemia) of some congolese plants”, Phytomedicine, 14, (2007), 192–195.

• E. Iyamu, E. Turner and T. Asakura, “In vitro effects of NIPRISAN (Nix-0699): a naturally occurring, potent antisickling agent” British Journal of Haematology, 118, 2002, 337–343J.

• O. Moody, O. O. Omotade, A. A. Adeyemo, “ Anti-sickling Potential of a Nigerian Herbal Formula (Ajawaron HF) and the Major Plant Component (Cissus populnea L. PK) ” Phytother Res. 17, (2003), 1173–1176.

• Gorecki et al., “Antisickling activity of amino acid benzyl esters” Proc. Natl. Acad. Sci.USABiochemistry ,1980,77(1), 181-185

• Iheanyichukwu elekwa, Michael monanu, Emmanuel anosike “In vitro effects of aqueous extracts of Zanthoxylum macrophylla roots on adenosine triphosphatases from human erythrocytes of different genotypes” BIOKEMISTRI 17, 1, 2005 :19-25.

• Stuart M.J., Nagel R.L., Sickle-cell disease. Lancet. 2004;364:1343-1360

• Steinberg M.H., Pathophysiologically based drug treatment of sickle cell disease. Trends in Pharmacological science. 2006;27:204-210

• Herfindol E.T., Gourley D.R., Text book of Therapeutics Drug and Disease Management.7th edi.,2001,251-254.

• Kate S. L., Lingojwar D. P., Epidemiology of Sickle Cell Disorder in the State of Maharashtra. Int J Hum Genet.2002;2:161-167

• Gupta R.B., Sickle cell disease loads in Madhypradesh. RMRCT update, 2006;3:1-8

• Balgir R.S., Epidemiology, Population Health Genetics and Phenotypic Diversity of Sickle Cell Disease in India. The Internet Journal of Biological Anthropology . 2007;1:2

Q&A…

Thank you for your attention

Sicklecare TAYUHM Limited.16, Adeyemi Lawson Street, Off Queens drive, Ikoyi Lagos, Nigeria.

+2348106910220.