Development of Piperine Derivatives for antiobesity effect

1. Development of Piperine Derivatives
For Anti-obesity effect
Dirgha Raj Joshi
College of Pharmacy
Medicinal Chemistry Lab
Wonkwang University

2. CONTENTS
INTRODUCTION
SYNTHETIC STRATEGIES
ACKNOWLEDGEMENT
CONCLUSION
RESULT AND DISCUSSION

3. BMI (Kg/m2) Classification
< 18.5 Under weight
18.5-24.99 Normal range
≥ 25 Over weight
≥ 30 Obese
International classification of obesity according to BMI
 What is Obesity?
 Accumulation of excess fat in different body parts
6 types of fat accumulation
 In Korea, 32.8% of adults are obese
 the men are 36.1% and women are 29.7%
 Different classification in Korea
 BMI ≥ 23kg/m2 is overweight and
 BMI ≥ 25 kg/m2 is obese
INTRODUCTION
3

4.  Main causes of obesity
 Imbalance between the intake of high energy food and
insufficient physical activities
 Other: genetics, medical cause, or psychiatric illness,
inadequate sleep, sophisticated lifestyle, endocrine
disruptors, some drugs side effect, pregnancy at later
age etc.
INTRODUCTION
 Complications with obesity
 Diabetes, hyperlipidemia etc.
4
 Management of obesity
 Pharmacological way: by using
drugs
 Non-pharmacological way:
Exercise, Yoga, Meditation, and
various Asanas
 Diet approach: controlling diet

5.  Management of obesity: Pharmacological
 Reducing food intake: Sibutramine,
withdrawn due to cardiovascular side effects
Sibutramine
Orlistat
 Modulating the central controller regulating
body weight: Lorcaserin, act on 5HT2c
serotonine receptor and reduces appetite
Ephedrine
 Modulating fat or protein
metabolism or storage:
leptin
 Increasing thermogenesis/
energy expenditure
Caffeine
 Reducing intestinal fat absorption by inhibiting
pancreatic lipase: Orlistat
INTRODUCTION
Lorcaserin
5

6. Drug Mechanism of action Avg. wt.
loss (%)* Adverse effects and comments
Orlistat
Pancreatic lipase inhibitor,
thereby reducing fat absorption
4
GI bloating and diarrhea, should be taken
with a multivitamin, to compensate for the
impaired absorption of fat soluble vitamins
Lorcaserin
Serotonin receptor agonist,
which acts in the brain to reduce
food intake
3
headache, dizziness, nausea, dry mouth and
constipation; avoid use with other serotonergic
drugs
Liraglutide
Glucagon-like receptor 1 agonist,
which reduces food intake
6
Nausea, vomiting, acute pancreatitis, gall bladder
disease, hypoglycemia
Diethylpropion,
phentermine,
phendimetrazine and
benzphetamine
Noradrenergic drug,
which functions as an appetite
suppressant
NA
Dizziness, dry mouth, insomnia, constipation,
irritability and cardio stimulatory effects
Phentermine–
topiramate
(extended release)
as appetite suppressants
through the release of serotonin,
noradrenaline and dopamine
9
paresthesia and change in taste (dysgeusia),
metabolic acidosis and glaucoma are rare; avoid
in MAOI antidepressant and in pregnancy
Naltrexone–
bupropion
(sustained release)
increase satiety and decrease
appetite, inhibiting the reuptake of
dopamine and noradrenaline,
blocking μ-opioid receptor and
activating pro-opiomelanocortin
6
Nausea, constipation and headache; avoid in
patients receiving opioids, MAOI antidepressants
and those with a history of seizures; the
cardiovascular safety of this treatment remains
uncertain
INTRODUCTION
 FDA approved drugs for obesity
6

7. Drugs
Years
on market
Side effect
Thyroid Extract 1892-present insulin suppression, heart complications, cardiac stress, hyperthyroidism
2,4 Dinitrophenol
(DNP)
1980s-
present
(without FDA
approval)
skin rash, cataracts, neuropathy, combination of hyperthermia, tachycardia,
diaphoresis, and tachypnea eventually leading to death, 62 deaths had been
attributed to DNP by 2011
Amphetamine 1947-1979
addiction, potential for abuse, alerted sexual behavior, cardiac arrhythmias,
ulcers, convulsions, coma, death
Phentermine 1959-present
fainting, swelling of feet or lower legs, trouble breathing, depression,
drowsiness, increased blood pressure, irritability, nervousness, blurred vision,
psychosis, skin rash, itching, stomach pain, unpleasant taste
Diethylpropion 1959-present
irregular heartbeat, heart palpitations, blurred vision, skin rash, itching, difficulty
breathing, chest pain, fainting, swelling of the ankles or feet
"Rainbow Pills“ 1940s-1960s rainbow pill" cocktails was linked to several deaths
Fenfluramine 1973-1997
withdrawn by the FDA on Sep. 15, 1997 because about 30% of patients had
abnormal echocardiograms . fatal lung or heart problems, difficulty sleeping,
irritability, vomiting, diarrhea, constipation
Dexfenfluramine 1973-1997
Withdrawn by the FDA on Sep. 15, 1997 due to 30% of patients had abnormal
echocardiograms. heart valve disease, abnormal heart rhythm, feeling faint,
drowsiness, heart throbbing
INTRODUCTION
 15 Anti-obesity drugs, 1892-present
7

8. Drugs
Years
on market
Side effects
Sibutramine 1997-2010
Withdrawn by the FDA on Oct. 8, 2010, increase risk of heart attack, stroke, and
cardiovascular death; abnormal liver function tests, widening of blood vessels,
throat and sinus irritation and congestion
Orlistat
1999-
present(Xenical)
2007-present
(Alli)
lowered absorption of vitamins, severe liver problems, kidney problems,
gallstones, oily rectal discharge, passing gas with oily discharge, urgent need to
have a bowel movement, oily or fatty stools, uncontrollable bowel movements
Hydroxycut
Diet
Supplements
2002-2009
jaundice, elevated liver enzymes, liver damage, seizures, CVS disorders,
rhabdomyolysis
Rimonabant
Rejected by the FDA
in June 2007
increased risk of seizures, depression, anxiety, insomnia, aggressiveness, and
suicidal thoughts; five deaths and 720 adverse reactions in the UK between
2005 and 2007
Lorcaserin
Approved by the FDA
on June 27, 2012
constipation, valvular heart disease, changes in attention or memory,
depression, suicidal thoughts, painful erections
Phentermine
& Topiramate
Approved by the FDA
on July 17, 2012
birth defects, depression, mood problems, insomnia, concentration, memory,
and speech difficulties, metabolic acidosis, low blood sugar in people with type
2 diabetes, seizures, kidney stones
Bupropion &
Naltrexone
In phase 3 clinical
trials, previously
rejected by the FDA
nausea, constipation, and vomiting in patients with type 2 diabetes; other side
effects unclear as of Apr. 25, 2014
INTRODUCTION
 15 Anti-obesity drugs, 1892-present
8

9. Piperine
 Discovered in 1819 by Hans Christian, from the fruits of Piper nigrum (The King of
species)
 Also found in P. longum and P. officinarum
 The amount of piperine varies from 1–2% in long pepper, to 5–10% in white and black
pepper
 Diverse ethnomedicinal practice, which were scientifically proven; antihypertensive,
antioxidant, antiplatelet, anti-obesity, antitumor, anticonvulsant, anti-thyroid, analgesic,
anti-inflammatory, antidiarrheal, antispasmodic, antidepressants, immunomodulatory,
antibacterial, antifungal, hepatoprotective, etc
 Piperine significantly reduces levels of plasma TC, LDL, VLDL, and activity of HMG CoA
reductase in liver, heart, and aorta, also significantly increase the level of plasma
lecithin cholesterol acyltransferase (LCAT), plasma and tissue lipoprotein lipase (LLP).
Piperine
C17H19NO3
285.34
Country Production
Vietnam 163
Indonesia 89
India 53
Brazil 42
China 31
World 473
Top black pepper producers in 2013
(thousands of tons)
INTRODUCTION
Piperine
long pepper
9

10.  Inhibits adipogenesis by antagonizing PPARγ activity as well as suppressing PPARγ
expression in 3T3-L1 Cells, therefore attenuates differentiation of fat cells
 Regulates hepatic lipid accumulation via alteration in the LXRα-mediated lipogenesis
INTRODUCTION
 Acutely toxic to mice, rats and hamsters
• The LD50 values for a single i.v., i.p., s.c., i.g. and i.m. administration
to adult male mice were 15.1, 43, 200, 330 and 400 mg/kg, respectively
 Most animals given a lethal dose died of respiratory paralysis within 3-17 min.
 In subacute toxicity studies, the rats died within 1-3 days after treatment
PPAR -alpha and -gamma pathways Biological role of LXR receptor Differentiation of 3T3L1 cells to adipocytes
Mechanism of action of piperine
Toxicity of piperine
10

11.  And curcumin in suppression of diethylnitrosamine (DENA)-induced hepatocellular
carcinoma (HCC) in rats
 And curcumin in modulating benzo(a)pyrene induced redox imbalance in mice lungs
 And paclitaxel on cell fate via cyt-c, Bax/Bcl-2-caspase-3 pathway in ovarian
adenocarcinomas SKOV-3 cells
 And ciprofloxacin on E. coli, Bacillus subtilis
 enhances the effect of many drugs
(including those metabolized by CYP3A4 and CYP2E1 enzymes):
 Diclofenac, Ibuprofen, Carbamazepine, Chlorzoxazone, Ampicillin trihydrate, Norfloxacin,
 Nevirapine, Domperidone, Docetaxel, Glimepiride, Nateglinide, Metformin, Fexofenadine
Paclitaxel
Curcumin Ciprofloxacin Carbamazepine
Synergistic effect of piperine
11
INTRODUCTION

12. Strategy for Derivatization:
SYNTHETIC STRATEGIES
12

13. Retrosynthetic analysis (2)
using NHS ester
with various amines
Retrosynthetic analysis (1)
using phosphonoamide
Retrosynthetic analysis
13
SYNTHETIC STRATEGIES

14. i) triethylphosphite, 120 °C, 12h. ii) p-methylthiobenzaldehyde, anhyd. LiOH, dry THF, 4Å MS, reflux, 24 h.
iii) LiOH. H2O, MeOH: H2O (10:1), THF, rt, 48 h. iv) various amines, CDI, dry DCM, rt, 12 h
 Key Steps
SYNTHETIC SCHEMEs
 Synthesis of piperine derivatives containing para-SMe
Horner-Wadsworth-Emmons (HWE) reaction Peptide bond formation
14

15. SYNTHETIC SCHEMEs
 Synthesis of piperine derivatives containing para-SMe
15

16. i) 1M KOH, water, 40 mins. ii) NHS, DCC, THF, rt, 24 h. iii) mono-Boc-piperazine, THF, 24 h.
iv) 5% TFA, DCM, rt, 16 h. v) tBuOK, THF, 0 °C, 6 h. vi) a) triphosgene, Na2CO3, TEA, dry hexane,
methanol (DR-216), (ethanol (DR-212), isopropanol (DR-214)).
SYNTHETIC SCHEMEs
 Synthesis of piperine derivatives containing para-SMe
16

17. SYNTHETIC STRATEGIES
Failed Overcome
DCM solvent used dry n- hexane used
piperazine reacted with triphosgene alcohol reacted with triphosgene
excess Na2CO3 and Hünig base used excess Na2CO3 and catalytic amount of TEA used
reaction done at 0.1 M dilution highly diluted and dropwise addition of reagents
Overcome
Failed
Solution of Synthetic problem in preparation of carbamate
from piperazine
17

18. i) Boc2O, Na2CO3, DCM/H2O, 22h. ii) MsCl, TEA, DCM, 0 °C, 2 h.
iii) DBU, DMF, 95 °C. iv) Conc. HCl, rt, 1 h. v) 7, TEA, DCM, rt.
vi) a) (DR-206) p-methylthiobenzaldehyde, tBuOK, DCM.
b) (DR-207) p-methoxythiobenzaldehyde.
Synthesis of piperine derivatives containing tetrahydropyridine
SYNTHETIC SCHEMEs
18

19.  Synthesis of piperine derivatives containing para-OMe
SYNTHETIC SCHEMEs
i) p-methoxybenzaldehyde, anhyd. LiOH, dry DCM, 4Å MS, reflux, 12 h. ii) anhyd. LiOH, MeOH, rt, 48 h.
iii) EDAC, NHS, dry DCM, rt, 20 h. iv) various amines, dry DCM, rt, 12 h. v) PDC, dry DCM, rt, 12 h.
19

20. i) p-methoxybenzaldehyde, tBuOK, THF, 0 °C. ii) a) triphosgene, Na2CO3, TEA, hexane
b) then, methanol (DR-215), ethanol (DR-211), isopropanol (DR-213), in case of R=Bn (DR-208),
cbz-Cl was used and ditertbutyldicarbonate (DR-279)
 Synthesis of piperine derivatives containing para-OMe
20
SYNTHETIC SCHEMEs

21.  Synthesis of piperine derivatives with one double bond
21
SYNTHETIC SCHEMEs

22. i) NHS, DCC, THF, rt, 24 h. ii) mono-Boc-piperazine, THF, 24 h. iii) 5% TFA, DCM, rt, 16 h.
iv) tBuOK, THF, 0 °C, 6 h. a) p-methoxy benzaldehyde, b) p-methylthio benzaldehyde, c) piperonal.
v) triphosgene, Na2CO3, TEA, hexane. a) methanol (DR-224-2, DR-225-2, DR-223-2),
b) cyclopentanol (DR-220, DR-221, DR-222), c) Boc2O (DR-226, DR-227, DR-228)).
Synthesis of piperine derivatives with one double bond
22
SYNTHETIC SCHEMEs

23.  Dimer derivatives
 dimer as a by-product (~ 10-40%),
collected from different carbamate formation reactions
23
SYNTHETIC SCHEMEs

24.  Synthesis of meta-paradioxy and dithiopiperine derivatives
i) dimethylthiocarbomyl chloride, 4% aq. NaOH, THF, 0 °C, 12h.
ii) 9, 27, tBuOK, dry DMF/ THF (1:3), 0 °C, 2h. iii) Boc2O, THF, 1.5h
iv) piperidine, EDAC, DCM, 4h. v) 27, tBuOK, dry DCM, 0 °C, 12h. vi) Ph2O, 220 °C, 10 h.
24
SYNTHETIC SCHEMEs

25.  3T3-L1 adipocytes cells
RESULTS AND DISCUSSION
25
Simvastatin

26. RESULTS AND DISCUSSION
 3T3-L1 adipocytes cells
26

27. RESULTS AND DISCUSSION
 3T3-L1 adipocytes cells
27

28. Simvastatin
RESULTS AND DISCUSSION
 3T3-L1 adipocytes cells
28

29. Diosgenin Simvastatin GW4064
RESULTS AND DISCUSSION
 HepG2 Cells
29

30. RESULTS AND DISCUSSION
 HepG2 Cells
30

31. RESULTS AND DISCUSSION
 HepG2 Cells
31

32. RESULTS AND DISCUSSION
 HepG2 Cells
32

33. CONCLUSION
 Future modification to the thio and to the
amide ring considering DR-049 could be
better
 p-Methylthio group gave better
activities over p-methoxy derivatives
 The higher lipophilic group attached
to the piperazine increased activity
and vice versa
 Monomer piperazine carboxylate better
over the dimer piperazine derivatives
 The 2 double bond aliphatic chain
prefer over 1 double bond
33

34. ACKNOWLEDGEMENT
 Supervisor:
Prof. Dr. Hak Sung Kim
 Examiner committee members:
Prof. Dr. Hyeon Cheol Oh
Prof. Dr. Youn-Chul Kim
 Dr. Soon-Ai Kim
 Wonkwang University, Graduate School
 Bioway Pharmaceutical Company
 All Seen and unseen helping hands