1 discovery of anti-leishmanial leads from natural lead h. e. j. research institute of chemistry...
TRANSCRIPT
![Page 1: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/1.jpg)
1
Discovery of Anti-Leishmanial Leads from Natural Lead
H. E. J. Research Institute of Chemistry
International Center for Chemical and Biological SciencesUniversity of Karachi, Karachi-75270, Pakistan
Sammer Yousuf and M. Iqbal Choudhary
04/21/23
![Page 2: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/2.jpg)
2
One of the greatest public health problems in many developing countries.
Second most prevailing tropical disease, after malaria.
Widely distributed in 88 countries of world including Afghanistan, Pakistan, Bangladesh, India, China and the southern United States and Europe.
Transmitted by the bite of infected sandfly (Phelobotomus).
Leishmaniasis-An Old Disease
04/21/23
![Page 3: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/3.jpg)
3
Leishmaniasis
WHO has estimated that more than 350 million people are at risk of infection with Leishmania species with 600,000 new clinical cases reported annually.
04/21/23
![Page 4: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/4.jpg)
4
Classification of Leishmaniasis
Caused by Lieshmania species in humans, depending on the ability of the organism to penetrate in the deep tissues.
• Visceral Leishmaniasis: Visceral leishmaniasis, the most severe clinical form, is caused by the Leishmania donovani. It characterized by its affects on internal organs, such as liver, spleen and bone marrow.
•Mucocutaneous Leishmaniasis: Mucocutaneous leishmaniasis can cause facial disfiguration due to erosion in the mucocutaneous sites of the mouth and
nose. 04/21/23
![Page 5: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/5.jpg)
5
•Cutaneous Diffuse Leishmaniasis: Cutaneous diffuse Leishmaniasis is characterized by the formation of nodules, multiple lumps, specially around the face and
on the external surface of arms and legs.
•Cutaneous Leishmaniasis: Cutaneous leishmaniasis is a least severe form of leishmaniasis. Leishmania maxicana and Leishmania major is responsible for cutaneous leishmaniasis.
Classification of Leishmaniasis
04/21/23
![Page 6: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/6.jpg)
6
Geographical Distribution of Cutaneous Leishmaniasis due to Leishmania major
04/21/23
90% of the cutaneous leishmaniasis cases are reported from Afghanistan, Brazil, Iran, Peru, Saudi Arabia and Syria
![Page 7: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/7.jpg)
7
Leishmaniasis in Pakistan
Major outbreaks were observed in NWFP, Jacobabad, Larkana, and Dadu districts of Sindh province.
Prevalent in Pakistan and has been reported from
almost all parts.
Diagnosis and treatment is difficult due to poor access to health services.
04/21/23
![Page 8: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/8.jpg)
8
• Leishmaniasis is a poverty-related disease. • It affects the poorest of the poor and is associated
with • Malnutrition • Weakness of the immune system • Lack of resources
According to WHO!
04/21/23
![Page 9: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/9.jpg)
9
• Antimonial compounds, amphotericin B, pentamidine, pyrazolopyrimidine, aminosidine, with a combination of antimony salts-pyrazolopyrimidine
• Stiff joints, gastrointestinal problems, cardiotoxicity and in some cases hepatic and renal insufficiency.
Current Treatment
04/21/23
![Page 10: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/10.jpg)
10
• Pakistan is a country with a rich tradition of medicinal herbs used by people to treat their health disorders
• Physalis angulata and Physalis purviana has already been reported previously for their leishmanicidal potential
Search Start
04/21/23
![Page 11: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/11.jpg)
11
•Physalis minima (Linn. var. indica) is a herb which is bitter in taste and widely used in folk medicines.
•The plant is used in vitiated conditions of gallbladder, burning sensation, bronchitis, for the treatment of inflammation, enlargement of spleen, urinary disorders, abdominal troubles and headache.
•The roots are used as a vermifuge and febrifuge.
•The 50% alcoholic extracts of the plant showed in vivo antimalarial, in vitro antigonorrhoeal activity.
BIOLOGICAL AND PHARMACOLOGICAL IMPORTANCE OF PHYSALIS MINIMA
04/21/23
![Page 12: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/12.jpg)
12
The fruits of Physalis minima are used as appetizer and have also been recorded as an ingredient of medicinal oil, which is given for spleen disorders.
• Antimycobacterial
• Immunomodulatory
• Hypoglycemic
BIOLOGICAL AND PHARMACOLOGICAL IMPORTANCE OF PHYSALIS MINIMA
04/21/23
![Page 13: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/13.jpg)
13
Methanolic Extract
Extracted with pet. ether (20 L)
(2.5 Kg)
Pet. ether Extract (200 g)
Insoluble part
Extracted with CH2Cl2 (20 L)
Chlorofrom Extract (150 g)
Ethylacetate Extract ( 200 g)
Extracted with Etylacetate (20 L)
Insoluble part
Insoluble part
Methanol water soluble part ( 200 g)
Extracted with MeOH: H2O (1:1)
Insoluble part
(20 L)
EXTRACTION OF EXTRACTION OF PHYSALIS MINIMAPHYSALIS MINIMA
04/21/23
![Page 14: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/14.jpg)
14
Medicinal Plant Plant’s Extracts (IC50 µg/mL ± S.D) % Reduction in
Amastigotes
(25 µg/mL ± S.D)
Physalis minima
Linn.
(Sun berry)
Methanol 26.00 ± 0.59 ND
Pet. Ether 31.62 ± 0.30 65
Chloroform 2.05 ± 0.02 90
Ethyl acetate 35.36 ± 0.08 ND
Standard Drugs Amphotericin B 0.12 ± 0.10 73.25 at 10µM
Pentamidine 5.13 ± 0.02 52.05 at 12.5µM
In Vitro leishmanicidal activity of Physalis minima extracts Against Promastigotes and Amstigotes Stages of
Leishmania major Parasite
04/21/23
![Page 15: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/15.jpg)
15
In vitro Cytotoxic Activity Against 3T3 Cell Lines (Normal Fibroblast cell).
Extracts of Physalis minima IC50 (µg/ mL) ± S.D.
3T3 cell lines
Methanol 18.78 ± 0.21
Pet. ether > 100
Dichloromethane 13.31 ± 0.70
Ethyl acetate >100
Standard
(Cyclohexamide)
0.26 ± 0.04
04/21/23
![Page 16: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/16.jpg)
16
Animal model Doses (mg/Kg) LD50 or Lethality
Wister Rats
(Male 200-250g)
10 Non Lethal
100 Non Lethal
500 Non Lethal
1000 Non Lethal
Animal Toxicity Determination (LD50) Against the
all Extracts of Physalis minima.
04/21/23
![Page 17: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/17.jpg)
17
In Vivo Leishmanicidal Studies on BALB/c Mice Model of Cutaneous Leishmanisis by
Topical Application of Gel having Extracts of Physalis minima Linn.
04/21/23
![Page 18: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/18.jpg)
18
Based on in vitro antileishmanial studies
Most active and least toxic mathanol and chloroform extracts were selected for
In vivo studiesLimited clinic trials
04/21/23
In Vivo Leishmanicidal Studies
![Page 19: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/19.jpg)
19
Development of Cutaneous Leishmania Lesion
First day of inoculation 5-days of infection inflammation showed
04/21/23
Inoculated in the base of the tail with 1x 106 to 5x 106
infective promastigotes
![Page 20: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/20.jpg)
20
Treatment of Infected Mice by Using White Soft Paraffin Based Gel Containing 25% Plant Extract
2-weeks of treatment Complete recovery after 4 weeks of
treatment.
04/21/23
1-weeks of treatment
![Page 21: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/21.jpg)
21
Clinical Trials For Cutaneous Leishmaniasis
By Using 25% Chloroform Extract of Ariel Parts of Physalis minima
at
Shaheed Mohtarma Benazir Bhutto Medical University, LARKANA
Conducted and Supervised by
PROF. Dr. Farooq Rahim Somroo04/21/23
![Page 22: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/22.jpg)
Before treatment After treatment
Topical Application of 25% Chloroform Extract of Ariel Parts of Physalis minima (Twice in a day)
04/21/23 22
![Page 23: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/23.jpg)
Before treatment After treatment
Topical Application of 25% Chloroform Extract of Ariel Parts of Physalis minima (Twice in a day)
04/21/23 23
![Page 24: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/24.jpg)
Before treatment After treatment
Topical Application of 25% Chloroform Extract of Ariel Parts of Physalis minima (Twice in a day)
04/21/23 24
![Page 25: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/25.jpg)
Topical Application of 25% Chloroform Extract of Ariel Parts of Physalis minima (Twice in a day)
54 Patients : Follow up for 2-4 weeks
39 Patients responded to treatment
39 Patients: Showed Recovery Response After Two Weeks of Treatment
39 Patients: Complete Recovery in 4 weeks
04/21/23 25
![Page 26: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/26.jpg)
26
Clinical Trials For Cutaneous Leishmaniasis
By Using 25% Methanolic Extract of Ariel Parts of Physalis minima
at
Shaheed Mohtarma Benazir Bhutto Medical University, LARKANA
Conducted and supervised by
PROF. Dr. Farooq Rahim Somroo04/21/23
![Page 27: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/27.jpg)
Before treatment After treatment
Topical Application of 25% Mathanolic Extract of Ariel Parts of Physalis minima (Twice in a day)
04/21/23 27
![Page 28: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/28.jpg)
Before treatment After treatment
Topical Application of 25% Mathanolic Extract of Ariel Parts of Physalis minima (Twice in a day)
04/21/23 28
![Page 29: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/29.jpg)
Before treatment After treatment
Topical Application of 25% Mathanolic Extract of Ariel Parts of Physalis minima (Twice in a day)
04/21/23 29
![Page 30: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/30.jpg)
Topical Application of 25% Methanolic Extract of Ariel Parts of Physalis minima (Twice in a day)
37 Patients : Follow up for 2-4 weeks
19 Patients responded to treatment
12 Patients: Showed Recovery Response After Two Weeks of Treatment
19 Patients: Complete Recovery in 4 weeks
04/21/23 30
![Page 31: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/31.jpg)
Chemical Constituents of Methanol and Chloroform Extracts of Physalis minima
Mathanol and Chloroform extracts are rich in
secondary metabolites commonly known as
steroidal lactones
Physalins
Withanolides
04/21/23 31
![Page 32: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/32.jpg)
32
PHYSALINS
CH3
O
CH3
O
HO
OO
O
OH
O
CH3
O1
3 5 7
9
19 11 13
14
16
1820
21
23 25
26
27
28
H
![Page 33: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/33.jpg)
33
CH2Cl2: MeOH 95: 5
CH2Cl2: MeOH 98: 2
CH2Cl2: MeOH 96: 4
CH2Cl2: MeOH 95: 5
CH2Cl2: MeOH 93: 7
CH2Cl2: MeOH 91: 9
Pet. ether: Acetone 80: 20
Pet. ether: Acetone 90: 10
Pet. ether: Acetone 84: 16
Pet. ether: Acetone 85: 15
CH2Cl2: MeOH 95: 6
Pet. ether: Acetone 80: 20
Dichloromethane Extract
Column Chroromatography (CC)
Fr. 1 (1.4 g)
Fr. A (120.5 mg)
Fr. B(98.8 mg)
Fr. E(88.5 mg)
5,6-Epoxy physalin B (3)Physalin H (1)
2,3-Dihydro-5,6-epoxy-3-methoxy-physalin B (6)
(25.2 mg)
(20.0 mg)
(12.2 mg)
(15.4 mg)
Isophysalin B(2)(20.2 mg)
27-Hydroxy-1-oxo-witha-24-enolide (8)
(20.0mg)
6-Deoxy physalin D (4)
(10.0 mg)
6-Oxo-11-hydroxyphysalin D (5)
(20.0 mg)
11-Hydroxyphysalin D (3)
Fr. C(50.5 mg)
6-Deoxy-11-hydroxyphysalin I (7)
(10.0mg)
Fr. 2 (2.5 g)
Physalin H(505 mg)
Fr. D(40.5 mg)
PURIFICATION OF PHYSALINS FROM PURIFICATION OF PHYSALINS FROM PHYSALIS MINIMA EXTRACTPHYSALIS MINIMA EXTRACT
![Page 34: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/34.jpg)
34
Antileishmanial Activity of Physalins
IC50 (M)= 0.92 0.001
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
OHOH
HO H
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
OH
H
IC50 (M)= 5.00 0.01
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
OH
H
O
HO
IC50 (M)= 3.65 0.070 11 -Hydroxy physalin D 6-Oxo-11 -Hydroxy physalin D
6-Deoxy physalin D
Amphotericine B0.12 0.105
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
Cl
H
OH Physalin H
IC50 (M)= 3.39 0.005
![Page 35: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/35.jpg)
35
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
OCH3
HHO
IC50 (M)= 4.86 0.0562
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
H
O
IC50 (M)= 3.39 0.005
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
O
H
H3CO
IC50 (M)= 19.4 0.18 2,3-Dihydro-5,6b-epoxy-3-methoxy-physalin B (4)
6-Dehydroxy-11-hydroxy-physalin I (7)
Amphotericine B0.12 0.105
O
O O
H
O
HCH3
OO
OHO
HO
CH3
H
CH3
H
Isophysalin B
IC50 (M)= 7.05 0.05 5,6-Epoxyphysalin B
![Page 36: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/36.jpg)
The encouraging results of this study
are suggestive that we are on the way
for developing cost effective/efficient
local therapy (Ointment) for the
treatment of cutaneous leishmaniasis
in the near future.
04/21/23 36
![Page 37: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/37.jpg)
37
CONCLUSION
Physalis minima was identified as potent antileishmanial agent against cutaneous leishmaniasis by employing in vitro, in vivo and limited clinical trails.
Methanol and Chloroform extracts of the plant were found to be the most active one therefore selected for the limited clinical trails
![Page 38: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/38.jpg)
38
CONCLUSION
Chloroform, the most active extract was further subjected to detailed phytochemical investigation to isolate many steroidal lactones
Discovery is patented
“Formulations Against Cutaneous Leishmaniasis”Atta-ur-Rahman, M. Iqbal Choudhary, Sammer Yousuf, Samreen, Farooq Rahman Soomro, Shahida Perveen(US patent 8,287,921 B1), Date of Patent Oct. 16, 2012
![Page 39: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/39.jpg)
39
ACKNOWLEDGEMENT
Prof. Atta-ur-Rahman N.I., H.I., S.I., T.I.
Miss Samreen
Higher Education Commission, Islamabad, Pakistan its financial support
![Page 40: 1 Discovery of Anti-Leishmanial Leads from Natural Lead H. E. J. Research Institute of Chemistry International Center for Chemical and Biological Sciences](https://reader035.vdocument.in/reader035/viewer/2022062409/56649dc95503460f94ac0176/html5/thumbnails/40.jpg)
4004/21/23
Thank you