By Yuqiong Xia2014

Formulation of Biotech Products

Pharmaceutical Biotechnology

Keywords

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Formulation, expicients, parenteral, solubility, anti-, adsorption, preservative, antioxidiant, osmotic pressure

Administration, IV, SC, IM, IP, Pulmonary, nasal Biodegradable microspheres, self-regulating mAb, immunoconjugate

生物制药工程 夏玉琼 西安电子科技大学

Outline

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Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery

生物制药工程 夏玉琼 西安电子科技大学

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Components in marketed formulations生物制药工程 夏玉琼 西安电子科技大学

Solubility enhancers

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Amino acids such as lysine and arginine Surfactants such as SDS

Solubility increase with higher arginine concentration

生物制药工程 夏玉琼 西安电子科技大学

Protein adsorption and aggregation

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Protein can be adsorbed to the interface like water/air, water/container wall and water/needle

Adsorbed proteins form aggregates and precipitate

生物制药工程 夏玉琼 西安电子科技大学

Anti-adsorption and anti-aggregation agents

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1% Albumin (白蛋白) Adsorb to surfaces instead of drugs

Surfactant Adsorb to the hydrophobic interfaces Turn the interface hydrophilic

生物制药工程 夏玉琼 西安电子科技大学

Buffer components

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Phosphate, citrate and acetate buffer Short, temporary pH change can cause

aggregation

http://3.imimg.com/data3/CA/NX/MY-5149559/buffer-solution-250x250.jpg

生物制药工程 夏玉琼 西安电子科技大学

Preservatives (防腐剂) and antioxidants

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Antioxidants belong to preservatives Easily oxidized amino acids Methionine, cysteine, tryptophan, tyrosine, histidine

Inert gas in the vials can reduce oxidation

生物制药工程 夏玉琼 西安电子科技大学

Antioxidants

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Ascorbic acid (抗坏血酸，维生素C)

Acetylcysteine （乙酰半胱氨酸）

http://www.medicalook.com/molecular/ascorbic_acid.gifhttp://ts1.mm.bing.net/th?id=H.4941541510548252&pid=1.7

生物制药工程 夏玉琼 西安电子科技大学

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Lyoprotectants Let proteins keep structures during freeze-drying Glycol

Cake formers Help to form tablet

Lyoprotectants/cake formers生物制药工程 夏玉琼 西安电子科技大学

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Freeze-drying

http://www.food-manufacturer.com/faqs/freeze-dried-knowledge/freeze-drying-process.html

Blue circles: Water molecules

dry dry rehydrate

生物制药工程 夏玉琼 西安电子科技大学

Osmotic agents

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Adjust the osmotic pressure Saline and mono- or disaccharide solutions

http://chemistry2.csudh.edu/rpendarvis/monosacch.html

生物制药工程 夏玉琼 西安电子科技大学

Outline

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Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery

生物制药工程 夏玉琼 西安电子科技大学

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Parenteral route

http://biology-forums.com/index.php?action=gallery;sa=view;id=8179

IM SC IV

生物制药工程 夏玉琼 西安电子科技大学

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Intraperitoneal (IP 腹腔) injections

http://ehs.uc.edu/lams/data/ratsmice/9040/40_030.html

生物制药工程 夏玉琼 西安电子科技大学

Blood half-life (t1/2) of drugs using parenteral route

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t-PA, a few minutes Monoclonal antibody (mAb), a few days IV Versus IM and SC Proteins using IV have shorter half-life Proteins using IV have more degradation reactions

and different deposition

生物制药工程 夏玉琼 西安电子科技大学

Oral route

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Patient friendly Low bioavailability Protein degradation in the gastrointestinal (GI) tract Poor permeability of the wall of the GI tract

http://www.uofmmedicalcenter.org/healthlibrary/Article/40233

生物制药工程 夏玉琼 西安电子科技大学

Oral route is suitable for vaccine

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To elicit an immune response, a little vaccine to target site is enough

生物制药工程 夏玉琼 西安电子科技大学

Nasal administration

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Only work for the nose Low bioavailability if systemic action is required

http://www.emsworld.com/article/10251608/intranasal-drug-administration

生物制药工程 夏玉琼 西安电子科技大学

Pulmonary（肺的）route

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Moderate bioavailability Patient friendly Fast uptake (faster than SC)

http://waleedtaha.tripod.com/rdds/

生物制药工程 夏玉琼 西安电子科技大学

Outline

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Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery

生物制药工程 夏玉琼 西安电子科技大学

Absorption enhancer in nasal administration

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Glycocholate (甘胆酸盐)

胰高血糖素

降血钙素

胰岛素

人生长激素

生物制药工程 夏玉琼 西安电子科技大学

Absorption enhancer in nasal administration

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Starch microsheres

1 empty starch microspheres2 soluble insulin 2.0 IU/kg3 soluble insulin IV 0.25 IU/kg4 insulin in starch microsphere 0.75 IU/kg5 insulin in starch microshphere 1.75 IU/kg

生物制药工程 夏玉琼 西安电子科技大学

Absorption enhancing using iontophoresis

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Transdermal iontophoretic delivery 离子电渗疗法

pH = 4 or 9

pH = 7.3

生物制药工程 夏玉琼 西安电子科技大学

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Absorption enhancing using iontophoresis

Yellow: IV 10 mg/kg; 0.025 mg/mlBlue: SC, 10 mg/kg; 0.025 mg/mlRed: iontophoresis, (1 mg/g; 0.17 mA/cm2; 5cm2 patch)

生物制药工程 夏玉琼 西安电子科技大学

Outline

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Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery

生物制药工程 夏玉琼 西安电子科技大学

PEGylation in rate-controlled delivery

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Polyoxyethylne glycol (PEG) attachment to proteins

PEG-UK has longer half-life

PEG-UK is more efficient

UK 尿激酶，治疗血栓

PT 凝血酶原时间，表征血栓

生物制药工程 夏玉琼 西安电子科技大学

Carrier system in rate-controlled delivery

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Open loop system Mechanical pumps Osmotically driven systems Biodegradable microspheres

Closed loop system Biosensor-pump combinations Self-regulating systems Encapsulated secretory cells

生物制药工程 夏玉琼 西安电子科技大学

Open loop systems: mechanical pumps

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Energy failure, problems with syringe, accidental needle withdrawal, leakage of catheter（导管） and problem with injection site

Problem with long-term drug stability The patient has to adapt to the pump rate http://www.cwladis.com/math104/bolus.jpg

生物制药工程 夏玉琼 西安电子科技大学

Osmotically driven systems

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Subcutaneously implantable, osmotic mini-pump Fixed rate

Semi-permeable membrane

Drug solution leaving via delivery portal

Flow moderator

Water entering

Drug reservoir

生物制药工程 夏玉琼 西安电子科技大学

Biodegradable microspheres

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PLGA （聚乳酸-聚羟基乙酸） microspheres prolonged the dosing interval of LHRH agonists from 1~3 months to 6 months

http://polyscitech.com/PLGA/PLGA.png

生物制药工程 夏玉琼 西安电子科技大学

Biodegradable microspheres

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Dextran（葡聚糖）-based microspheres

http://0.tqn.com/d/chemistry/1/0/s/E/1/Dextran.jpg

Dextran microspheres prolonged the release time of IgG from 10~ days to 20+ days

生物制药工程 夏玉琼 西安电子科技大学

Closed loop systems: biosensor-pump combinations

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A biosensor Measure the plasma level of the protein

An algorithm Calculate the required input rate

A pump system Administrate the drug at the required rate

生物制药工程 夏玉琼 西安电子科技大学

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Biosensor-pump combinations生物制药工程 夏玉琼 西安电子科技大学

Self-regulating systems

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Insulin release as a function of glucose concentration By competitive desorption By enzyme-substrate reaction

生物制药工程 夏玉琼 西安电子科技大学

By competitive desorption

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Con A with glycosylated insulin is administrated

ConA can bind to glucose more stronger than glycosylated insulin

生物制药工程 夏玉琼 西安电子科技大学

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Results of the above insulin delivery

Dogs with pancreatic problems

▲Dogs with pancreatic problems after insulin delivery○Normal dogs

生物制药工程 夏玉琼 西安电子科技大学

By Enzyme-substrate reactions

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Acid sensitive polymers, enzyme and insulin Enzyme catalyzes glucose to gluconic acid pH becomes low Acid-sensitive polymers release insulin

http://ts1.mm.bing.net/th?id=H.4516021944648358&pid=1.7http://ts4.mm.bing.net/th?id=H.4696088444078218&pid=1.7

enzyme

Glucose Gluconic acid

生物制药工程 夏玉琼 西安电子科技大学

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Encapsulated secretory cells (implanted)生物制药工程 夏玉琼 西安电子科技大学

Outline

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Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery

生物制药工程 夏玉琼 西安电子科技大学

Site-specific delivery

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Maximize therapeutic effect Avoid toxic effects

http://www.pharmainfo.net/santosh-kumar-jh/site-specific-targetting

生物制药工程 夏玉琼 西安电子科技大学

Carrier system in site-specific delivery

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Passive targeting Particles up to 100 nm can enter leaky tumor tissues,

but not compact normal tissues

Active targeting An active moiety A carrier A homing device

生物制药工程 夏玉琼 西安电子科技大学

“natural targeting devices”: antibody

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Fc activates complement Complement causes lysis of target cells

生物制药工程 夏玉琼 西安电子科技大学

Murine mAb as carrier

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Produce human anti-mouse antibody (HAMA) To avoid HAMA Use F(ab)2 or F(ab) fragments Use human Fc part and murine Fab part Use completely human MAb in transgenic mouse

生物制药工程 夏玉琼 西安电子科技大学

Bispecific antibodies

46http://www.whatisbiotechnology.org/assets/exhibitions/milstein/milstein-clinical-m5.png

Bispecific antibody brings target cells or tissuets to effector cells and trigger cytotoxity

Target cells or tissues Bind to effector cells

Bispecific antibody

生物制药工程 夏玉琼 西安电子科技大学

Immunoconjugates

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Conjugate drug to antibody

D: drugs

生物制药工程 夏玉琼 西安电子科技大学

Summary

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Excipients in parenteral formulation Routes of administration: IV, IM, SC, IP, oral, nasal,

Pulmonary Absorption enhancement in nasal route Carrier system in rate-controlled delivery:

PEGylation, biodegradable microshpere Carrier system in site-specific delivery: mAb,

biospecific antibody, immunoconjugate

生物制药工程 夏玉琼 西安电子科技大学

Homework 3 (choose one)

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Classify the excipients in Insulin Aspart Injection: glycerol, phenol, ZnCl2, Na2HPO4, NaCl, HCl/NaOH, steril water

Classify the excipients in EPO (促红细胞生成素) injection: human albumin, NaCl, sodium citriate, trisodium citrate

生物制药工程 夏玉琼 西安电子科技大学