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DMPK, Preformulation, Animal House, Genotox and Toxicology

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DMPK Service Offerings

AbsorptionPermeabilityCaco-2 (A B and B A);

PAMPA; MDCK-MDR-1

SolubilityAqueous (various pH);

SGF; SIF (kinetic and TD)

Pharmacokinetics in Mice (SAM, Nude mice;

Balb/c); Rats (S.D, Wistar);Rabbits and Dogs (outsourced)

Bile duct cannulated rat PK Cannulation of portal vein,

lymphatic vessel and urinarybladder in rats

Cassette dosing is possible Microsampling in mice Dose escalation studies DBS (dried blood spot)

DistributionProtein binding Equilibrium dialysis (ED);

Ultra filtrationTissue distribution in Rats (using cold compound)

ED method to determine fuin plasma, tumor, CSF and brain homogenate

ExcretionMass balance (using metabolic cages) Biliary and Urinary excretion

Cross functional activitiesPK-PD (with in vivo pharmacology)Toxicokinetics (with Toxicology)D2M prediction (with Development group)

MetabolismMetabolic stability Liver microsomes; S-9

fractions and Hepatocytes

CYP and FMO profilingCYP induction (PXR/in vivo)

CYP inhibition

Metabolite ID in vitro using liver

microsomes, hepatocytes in vivo from plasma, bile,

urine and feces

Glutathione trapping

Blood/plasma partitioning

Time dependent inhibition

Plasma and Chemical stability

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In vivo Pharmacokinetics

Rodent Dog Monkey

Species

Mice (C57BL/6, BALB/c, Nude, SCID, SAM) andRat (Sprague Dawley,

Wistar)

Beagle dog (tie-up with PalamurBio,

Mahaboobnagar, Hyderabad and Vimta Labs, Hyderabad)

Cynomolgus (Southern Research Institute, USA

and Covance, China)

Animal SourceVivo Bio Tech, Hyderabad;

Reliance, Mumbai; Harlan and Charles River

Breeding and colony maintained at PalamurBio by Isoquimen, Spain.

Details will be provided on request

Routes of administration

Oral, Intravenous, Subcutaneous, Intraperitoneal, Perfusion etc. Capability to cannulate bile duct, portal vein, urinary bladder etc.

Blood sampling Retro-orbital /Tail vein / Jugular Vein (100 µL) Jugular Vein and Radial Vein (0.5 to 1 mL)

Regimen Single or Repeated (4/7/14/28-day) dosing

Time points 0.08 or 0.17 (IV only), 0.25, 0.5, 1, 2, 4, 8, 10 (PO only) and 24 h

Matrix for analysis Plasma in case of PK studies (in rodents, dog and monkey). In case of rodents we can analyze NCE concentration in all tissues including brain

Turnaround time 5-6 days (PO + IV) 7 days (per route) Will be provided on request

Current capacity 20 compounds/week Will be provided on request

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Instrumentation and Software

Six LC-MS/MS instruments(API-6500, API-5500, API-4000: 2; Thermo Ultra and API-4000 Q-Trap with UPLC)

Three HPLC units Nitrogen evaporators Refrigerated centrifuges Deep freezers (-80oC and -20oC) Freeze-drier Tissue homogenizer Franz Diffusion cell Shaking water bath (Julabo) CO2 incubator Genevac evaporator (to remove DMSO) Microbalances with printer Positive pressure SPE Vacuum manifolds with pump Ultrasonicator Milli-Q water system UPS and Standby Generator

Franz diffusion cells – Skin permeation

RapidFire 365 – For high throughput screening

WinNonlin

Graphpad Prism

Galileo LIMS

LightSight software for MetID

Automaton (method development)

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Turnaround time for various DMPK studies

S.No Study title Turn around time1 Solubility in PBS, SGF, FaSSIF, FeSSIF 5-6 days

2 Metabolic stability in microsomes 5-6 days

3 CYP liability 5-6 days

4 Protein binding 5-6 days

5 Caco-2/MDCK-MDR-1/PAMPA assay 6-7 days

6 Stability in plasma/Chemical stability 5-6 days

7 Reactive metabolite study 5-6 days

8 In vivo PK in rodents 6-7 days

9 Tissue distribution 10 days

10 Biliary excretion 10 days

11 Bioanalytical method validation 15 days

12 Met-ID 15 days

13 CYP profiling 5-6 days

14 Time dependent inhibition 5-6 days

5

Throughput of studies in 2018

1917

776952

604918

230325

122

9

37

8600

1

10

100

1000

10000

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Expertise in bioanalysis

• Strong bioanalytical team with capabilities to enable fit-for-purposeand/or full validated HPLC and LC-MS/MS methods

• Wide range of in vitro and in vivo assays• Expertise in handling various matrices (plasma, blood, bile, urine, feces

and various tissues)• Array of sampling processing techniques like precipitation, liquid-liquid

extraction, solid-phase extraction and derivatization process• Enabled highly sensitive methods (pg/mL as LLOQ) for several client

programs• Developed fit-for-purpose bioanalytical methods for various biomarkers

using LC-MS/MS and RapidFire (to support efficacy / target engagementstudies)

• Chiral separations• Cassette analysis• Prodrug analysis

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Preformulation capabilities

pH Solubility profile in buffer

system

Solubility in biorelevant

media

Solubility in presence of

solubilizers

Crystal shape or habit

Particle size distribution

Bulk density and tap density

Thermal properties

Powder X-ray diffraction

Hygroscopicity

Solution state

Solid state

Solubility study Solid state characterization Stability study

Salt and polymorph screening and characterization

Selection of one lead and one back up for stable salt and polymorph

Characterization of salt and polymorph

Stability study

Solubility study

PK study in rodents or other species

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Preformulation capabilities (2 of 2)

Dissolution study

Intrinsic dissolution in

aqueous media

Dissolution at different pH

Dissolution in biorelevant

media

Bulk properties

Bulk density

Tap density

Compressibility and Housner ratio

Flow properties

Selection and recommendation of preferred

excipients for preclinical/ clinical dosage

form developmentExcipients compatibility study

Excipients compatibility study will be preformed with excipients, based on the intended dosage

form and formulation development strategy

Excipients compatibility study will be done by isothermal stress method and will be evaluated for

related substance by HPLC analysis

Based on excipients compatibility, excipients will be selected for further development work

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Preclinical formulations development-Oral Route

Drug/physicochemical/ study purpose/dose, animal species

Suspension (e.g: 0.5% methyl cellulose)

Micronized suspension

No

Yes

Yes

Solution (e.g.: water, pH buffer)

NoNo

Co-solvent and pH Cyclodextrins and pH

Micelles and pH

No

Nanosuspension

Solid dispersion

Emulsion, microemulsions, SMEDDS, solid lipid

nanoparticles

No

Discovery/ structural modification

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Preclinical formulations development- IV route

Drug/physicochemical/study purpose/ route, animal species

Solution (e.g: normal saline, D5W)

No

pH adjustmentCo-solvent and pH

Cyclodextrins and pHSurfactant and pH

No

Nanosuspension

Microemulsions, liposomes etc.

No

Discovery/structural modification

Yes

YesIn vitro serial dilution

evaluation for precipitation

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Jubilant Biosys is committed to the humane treatment to the research animals.Animals are treated with the highest standards of respect, compassion, andparticular attention is given to housing conditions, social interaction and enrichment.

The Animal Facility is registered with:“Committee for the Purpose of Control and Supervision ofExperiments on Animals” (CPCSEA) Reg. no. 1026/PO/RcBi/S/07/CPCSEA.(Ministry of Environment, Forest and Climate Change, Govt. of India)

Full Accreditation since June 2012 NGCMA certified facility (Jun 2015)

IAEC (Institutional Animal Ethics Committee) monitors activities related to animalfacility & ensure that all animal experimentation procedures are carried out strictly asper the guidelines and approved study protocols

Laboratory Animal Facility - Regulatory and Ethical Status

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Toxicology Capabilities

General toxicology (GLP/non-GLP)

• Maximum Tolerated Dose (MTD) study

• Dose range finding study (4, 7 and 14 days)

• Repeated dose toxicity studies (28 and 90 days) with Functional Observations Battery

• Toxicokinetics studies

Genotoxicology (GLP/non-GLP)

• Ames test / Mini-Ames study

• Micronucleus test (In vivo & In vitro)

• Chromosomal aberration test (In vitro)