Formulation of Biotech Products

Transcription

1 Pharmaceutical Biotechnology Formulation of Biotech Products By Yuqiong Xia 2014

2 Keywords Formulation, expicients, parenteral, solubility, anti-, adsorption, preservative, antioxidiant, osmotic pressure Administration, IV, SC, IM, IP, Pulmonary, nasal Biodegradable microspheres, self-regulating mab, immunoconjugate 2

3 Outline Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery 3

4 Components in marketed formulations 4

5 Solubility enhancers Amino acids such as lysine and arginine Surfactants such as SDS Solubility increase with higher arginine concentration 5

6 Protein adsorption and aggregation Protein can be adsorbed to the interface like water/air, water/container wall and water/needle Adsorbed proteins form aggregates and precipitate 6

7 Anti-adsorption and anti-aggregation agents 1% Albumin ( 白蛋白 ) Adsorb to surfaces instead of drugs Surfactant Adsorb to the hydrophobic interfaces Turn the interface hydrophilic 7

8 Buffer components Phosphate, citrate and acetate buffer Short, temporary ph change can cause aggregation 8

9 Preservatives ( 防腐剂 ) and antioxidants Antioxidants belong to preservatives Easily oxidized amino acids Methionine, cysteine, tryptophan, tyrosine, histidine Inert gas in the vials can reduce oxidation 9

10 Antioxidants Ascorbic acid ( 抗坏血酸, 维生素 C) Acetylcysteine ( 乙酰半胱氨酸 )

11 Lyoprotectants/cake formers Lyoprotectants Let proteins keep structures during freeze-drying Glycol Cake formers Help to form tablet 11

12 Freeze-drying dry dry rehydrate Blue circles: Water molecules 12

13 Osmotic agents Adjust the osmotic pressure Saline and mono- or disaccharide solutions 13

14 Outline Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery 14

15 Parenteral route IM SC IV 15

16 Intraperitoneal (IP 腹腔 ) injections 16

17 Blood half-life (t 1/2 ) of drugs using parenteral 西安电子科技大学 route t-pa, a few minutes Monoclonal antibody (mab), a few days IV Versus IM and SC 生物制药工程 Proteins using IV have shorter half-life Proteins using IV have more degradation reactions and different deposition 夏玉琼 17

18 Oral route Patient friendly Low bioavailability Protein degradation in the gastrointestinal (GI) tract Poor permeability of the wall of the GI tract 18

19 Oral route is suitable for vaccine To elicit an immune response, a little vaccine to target site is enough 19

20 Nasal administration Only work for the nose Low bioavailability if systemic action is required 20

21 Pulmonary( 肺的 )route Moderate bioavailability Patient friendly Fast uptake (faster than SC) 21

22 Outline Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery 22

23 Absorption enhancer in nasal administration Glycocholate ( 甘胆酸盐 ) 胰高血糖素降血钙素胰岛素人生长激素 23

24 Absorption enhancer in nasal administration Starch microsheres 1 empty starch microspheres 2 soluble insulin 2.0 IU/kg 3 soluble insulin IV 0.25 IU/kg 4 insulin in starch microsphere 0.75 IU/kg 5 insulin in starch microshphere 1.75 IU/kg 24

25 Absorption enhancing using iontophoresis Transdermal iontophoretic delivery 离子电渗疗法 ph = 4 or 9 ph =

26 Absorption enhancing using iontophoresis 26 Yellow: IV 10 mg/kg; mg/ml Blue: SC, 10 mg/kg; mg/ml Red: iontophoresis, (1 mg/g; 0.17 ma/cm 2 ; 5cm 2 patch)

27 Outline Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery 27

28 PEGylation in rate-controlled delivery Polyoxyethylne glycol (PEG) attachment to proteins UK 尿激酶, 治疗血栓 PT 凝血酶原时间, 表征血栓 PEG-UK has longer half-life PEG-UK is more efficient 28

29 Carrier system in rate-controlled delivery Open loop system Mechanical pumps Osmotically driven systems Biodegradable microspheres Closed loop system Biosensor-pump combinations Self-regulating systems Encapsulated secretory cells 29

30 Open loop systems: mechanical pumps 30 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

31 Osmotically driven systems Subcutaneously implantable, osmotic mini-pump Fixed rate Drug solution leaving via delivery portal Flow moderator Semi-permeable membrane 31 Water entering Drug reservoir

32 Biodegradable microspheres PLGA ( 聚乳酸 - 聚羟基乙酸 ) microspheres prolonged the dosing interval of LHRH agonists from 1~3 months to 6 months 32

33 Biodegradable microspheres Dextran( 葡聚糖 )-based microspheres Dextran microspheres prolonged the release time of IgG from 10~ days to 20+ days 33

34 Closed loop systems: biosensor-pump 西安电子科技大学 combinations 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 34

35 Biosensor-pump combinations 35

36 Self-regulating systems Insulin release as a function of glucose concentration By competitive desorption By enzyme-substrate reaction 36

37 By competitive desorption Con A with glycosylated insulin is administrated ConA can bind to glucose more stronger than glycosylated insulin 37

38 Results of the above insulin delivery Dogs with pancreatic problems Dogs with pancreatic problems after insulin delivery Normal dogs 38

39 By Enzyme-substrate reactions enzyme Glucose Gluconic acid Acid sensitive polymers, enzyme and insulin Enzyme catalyzes glucose to gluconic acid ph becomes low Acid-sensitive polymers release insulin

40 Encapsulated secretory cells (implanted) 40

41 Outline Excipients in parenteral formulations Delivery of proteins Routes of administration Absorption enhancement Carrier system in rate-controlled delivery Carrier system in site-specific delivery 41

42 Site-specific delivery Maximize therapeutic effect Avoid toxic effects 42

43 Carrier system in site-specific delivery 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 43

44 natural targeting devices : antibody Fc activates complement Complement causes lysis of target cells 44

45 Murine mab as carrier 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 45

46 Bispecific antibodies Bispecific antibody Target cells or tissues Bind to effector cells Bispecific antibody brings target cells or tissuets to effector cells and trigger cytotoxity 46

47 Immunoconjugates Conjugate drug to antibody D: drugs 47

48 Summary 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 48

49 Homework 3 (choose one) Classify the excipients in Insulin Aspart Injection: glycerol, phenol, ZnCl 2, Na 2 HPO 4, 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 49