生物技术药物制剂
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Dark, opaque walls reduce oxidation Air-tight containers or argon atmosphere
reduces air oxidation
Storage - Additives
Addition of stabilizing salts or ions (Zn+ for insulin)
Identified through high-throughput screens of >10,000 randomly generated compounds
Function predominantly intracellularly
Low specificity for targets
PEG is a non-toxic, hydrophilic, FDA approved, uncharged polymer
Increases in vivo half life (4-400X) Decreases immunogenicity 降低免疫原性 Improved stability towards proteolysis Increase solubility of the protein in aqueous
Storage - Packaging
Smooth glass walls best to reduce adsorption or precipitation
Avoid polystyrene聚苯乙烯 or containers with silanyl硅烷 or plasticizer coatings
Biotechnology is often used to refer to genetic engineering technology of the 21st century.
Biopharmaceuticals: current status
Approximately one in every three new drugs now coming on the market is a biopharmaceutical.
Biopharmaceuticals Biological macromolecules
produced by living systems Created from amino acids and
nucleic acids Rely on defined biological
pathways Function predominantly
How to Deal with These Problems?
Storage
Formulation
Delivery
Pharmaceutics
Storage – Refrigeration 冷藏
Low temperature reduces microbial growth and metabolism
enzymes)
Comparing Traditional Medical Discovery with Biotechnology
Traditional Medicines
Synthetic small molecules produced through chemical processes
extracellularly Have a targeted mechanism of
action
生物技术药物的特点
大多数是蛋白质或多肽类药物 ①药理活性高，剂量低； ②结构复杂，理化性质不稳定； ③口服给药易受胃肠道pH、菌群及酶系统破坏；
Most are administered by injection rather than via the oral route. ④生物半衰期短，体内清除率高； ⑤具有多功能性，作用广泛,免疫原性的问题 ； ⑥检测存在诸多困难和不便。
Protein Pharmaceutics
SБайду номын сангаасorage
Formulation
Delivery
Protein Formulation
Protein sequence modification 蛋白序列的修饰 (site directed mutagenisis 点突变)
PEGylation PEG化 Proteinylation 蛋白质化 Microsphere/Nanosphere encapsulation 微球或
纳米球包裹
Site Directed Mutagenesis
Allows amino acid substitutions at specific sites in a protein
i.e. substituting a Methionine甲硫氨酸 to a Leucine亮氨酸 will reduce likelihood of oxidation
very quickly Unwanted allergic reactions 过敏反应 may develop
(even toxicity) Loss due to insolubility/adsorption
The Problem with Proteins (in vitro - in the bottle)
Biotechnology-derived Products
Chapter 17 Delivery of Biopharmaceuticals
课程要求
掌握生物技术药物的概念、特点；熟悉多 肽蛋白质类药物注射剂的处方设计、制备 工艺与质量评价。
了解多肽蛋白质类药物的其它非注射剂的 类型；了解基因传递系统的概念、类型。
Introduction
生物技术药物（ Biopharmaceuticals, Biotechnology-derived products）是通过细 胞融合、重组DNA或转基因等技术，利用 细菌、酵母、昆虫、植物或哺乳动物细胞 等各种表达系统制备而成的，用于预防、 诊断或治疗疾病的药物。
Biopharmaceuticals are medical drugs produced using biotechnology.
Polypeptides composed of covalently linked amino acids
Polypeptides with <40 amino acids are called peptides
Polypeptides with >40 amino acids are called proteins
conditions Hard to obtain in large quantities Easily destroyed/eliminated by the body
The Problem with Proteins (in vivo)
Elimination清除 by B and T cells Proteolysis by peptidases 在肽酶的作用下水解 Small proteins (<30 kD) filtered out by the kidneys
Biotechnology 生物技术
Any technological application that uses biological systems, living organisms, or derivatives, to make or modify products or processes for specific use.
Low temperature reduces thermal or spontaneous denaturation
Low temperature reduces adsorption Freezing is best for long-term storage Freeze/Thaw can denature proteins
Over 200 biopharmaceutical products had gained marketing approval in the USA and/or EU.
All but three are protein-based therapeutic agents.
Classification of biopharmaceuticals
The Problem with Proteins
Very large and unstable molecules Structure is held together by weak noncovalent
bonds 非共价键 Easily destroyed by relatively mild storage
Addition of polyols (glycerol and/or polyethylene glycol) to solubilize
Addition of sugars or dextran to displace water or reduce microbe growth
Use of surfactants (CHAPS) to reduce adsorption and aggregation
Noncovalent
Denaturation 变性 Aggregation 聚集 Precipitation 沉淀 Adsorption 吸附
Covalent
Deamidation 脱酰胺 Oxidation 氧化 Disulfide exchange 二硫键交换 Proteolysis 蛋白水解
gonadotrophins) Haematopoietic growth factors 造血生长因子(Erythropoietin,
colony stimulating factors) Interferons 干扰素(Interferons-α, -β, -γ) Interleukin-based products 白介素类(Interleukin-2) Vaccines 疫苗(Hepatitis B surface antigen) Monoclonal antibodies 单克隆抗体 Additional products (tumor necrosis factor, therapeutic
Delivery of Biopharmaceuticals
Protein/peptide delivery Vaccine delivery Delivery of nucleic acid-based therapeutics
Protein/peptide delivery
Proteins
Freeze Drying
Freeze liquid sample in container Place under strong vacuum Solvent sublimates leaving only solid or
nonvolatile compounds Reduces moisture content to <0.1%
Function of a protein determined by its noncovalent 3D structure
Amino Acids
H3N+ H
O
O R
Polypeptides
H3N+ H
OH N
R1 H
R2 H N
OH
OH N
R3 H
R4 O
O
Levels of Protein Structure
Blood factors 血液因子(Factor VIII and Factor IX) Thrombolytic agents 溶栓类(tissue plasminogen activator) Hormones 激素类(insulin, glucagon, growth hormone,
Strategic placement of cysteines半胱氨酸 to produce disulfides
PEGylation
+
CH-CH-CH-CH-CH-CH-CH-CH-CH-CH || ||| || ||| OH OH OH OH OH OH OH OH OH OH
PEGylation
reduces air oxidation
Storage - Additives
Addition of stabilizing salts or ions (Zn+ for insulin)
Identified through high-throughput screens of >10,000 randomly generated compounds
Function predominantly intracellularly
Low specificity for targets
PEG is a non-toxic, hydrophilic, FDA approved, uncharged polymer
Increases in vivo half life (4-400X) Decreases immunogenicity 降低免疫原性 Improved stability towards proteolysis Increase solubility of the protein in aqueous
Storage - Packaging
Smooth glass walls best to reduce adsorption or precipitation
Avoid polystyrene聚苯乙烯 or containers with silanyl硅烷 or plasticizer coatings
Biotechnology is often used to refer to genetic engineering technology of the 21st century.
Biopharmaceuticals: current status
Approximately one in every three new drugs now coming on the market is a biopharmaceutical.
Biopharmaceuticals Biological macromolecules
produced by living systems Created from amino acids and
nucleic acids Rely on defined biological
pathways Function predominantly
How to Deal with These Problems?
Storage
Formulation
Delivery
Pharmaceutics
Storage – Refrigeration 冷藏
Low temperature reduces microbial growth and metabolism
enzymes)
Comparing Traditional Medical Discovery with Biotechnology
Traditional Medicines
Synthetic small molecules produced through chemical processes
extracellularly Have a targeted mechanism of
action
生物技术药物的特点
大多数是蛋白质或多肽类药物 ①药理活性高，剂量低； ②结构复杂，理化性质不稳定； ③口服给药易受胃肠道pH、菌群及酶系统破坏；
Most are administered by injection rather than via the oral route. ④生物半衰期短，体内清除率高； ⑤具有多功能性，作用广泛,免疫原性的问题 ； ⑥检测存在诸多困难和不便。
Protein Pharmaceutics
SБайду номын сангаасorage
Formulation
Delivery
Protein Formulation
Protein sequence modification 蛋白序列的修饰 (site directed mutagenisis 点突变)
PEGylation PEG化 Proteinylation 蛋白质化 Microsphere/Nanosphere encapsulation 微球或
纳米球包裹
Site Directed Mutagenesis
Allows amino acid substitutions at specific sites in a protein
i.e. substituting a Methionine甲硫氨酸 to a Leucine亮氨酸 will reduce likelihood of oxidation
very quickly Unwanted allergic reactions 过敏反应 may develop
(even toxicity) Loss due to insolubility/adsorption
The Problem with Proteins (in vitro - in the bottle)
Biotechnology-derived Products
Chapter 17 Delivery of Biopharmaceuticals
课程要求
掌握生物技术药物的概念、特点；熟悉多 肽蛋白质类药物注射剂的处方设计、制备 工艺与质量评价。
了解多肽蛋白质类药物的其它非注射剂的 类型；了解基因传递系统的概念、类型。
Introduction
生物技术药物（ Biopharmaceuticals, Biotechnology-derived products）是通过细 胞融合、重组DNA或转基因等技术，利用 细菌、酵母、昆虫、植物或哺乳动物细胞 等各种表达系统制备而成的，用于预防、 诊断或治疗疾病的药物。
Biopharmaceuticals are medical drugs produced using biotechnology.
Polypeptides composed of covalently linked amino acids
Polypeptides with <40 amino acids are called peptides
Polypeptides with >40 amino acids are called proteins
conditions Hard to obtain in large quantities Easily destroyed/eliminated by the body
The Problem with Proteins (in vivo)
Elimination清除 by B and T cells Proteolysis by peptidases 在肽酶的作用下水解 Small proteins (<30 kD) filtered out by the kidneys
Biotechnology 生物技术
Any technological application that uses biological systems, living organisms, or derivatives, to make or modify products or processes for specific use.
Low temperature reduces thermal or spontaneous denaturation
Low temperature reduces adsorption Freezing is best for long-term storage Freeze/Thaw can denature proteins
Over 200 biopharmaceutical products had gained marketing approval in the USA and/or EU.
All but three are protein-based therapeutic agents.
Classification of biopharmaceuticals
The Problem with Proteins
Very large and unstable molecules Structure is held together by weak noncovalent
bonds 非共价键 Easily destroyed by relatively mild storage
Addition of polyols (glycerol and/or polyethylene glycol) to solubilize
Addition of sugars or dextran to displace water or reduce microbe growth
Use of surfactants (CHAPS) to reduce adsorption and aggregation
Noncovalent
Denaturation 变性 Aggregation 聚集 Precipitation 沉淀 Adsorption 吸附
Covalent
Deamidation 脱酰胺 Oxidation 氧化 Disulfide exchange 二硫键交换 Proteolysis 蛋白水解
gonadotrophins) Haematopoietic growth factors 造血生长因子(Erythropoietin,
colony stimulating factors) Interferons 干扰素(Interferons-α, -β, -γ) Interleukin-based products 白介素类(Interleukin-2) Vaccines 疫苗(Hepatitis B surface antigen) Monoclonal antibodies 单克隆抗体 Additional products (tumor necrosis factor, therapeutic
Delivery of Biopharmaceuticals
Protein/peptide delivery Vaccine delivery Delivery of nucleic acid-based therapeutics
Protein/peptide delivery
Proteins
Freeze Drying
Freeze liquid sample in container Place under strong vacuum Solvent sublimates leaving only solid or
nonvolatile compounds Reduces moisture content to <0.1%
Function of a protein determined by its noncovalent 3D structure
Amino Acids
H3N+ H
O
O R
Polypeptides
H3N+ H
OH N
R1 H
R2 H N
OH
OH N
R3 H
R4 O
O
Levels of Protein Structure
Blood factors 血液因子(Factor VIII and Factor IX) Thrombolytic agents 溶栓类(tissue plasminogen activator) Hormones 激素类(insulin, glucagon, growth hormone,
Strategic placement of cysteines半胱氨酸 to produce disulfides
PEGylation
+
CH-CH-CH-CH-CH-CH-CH-CH-CH-CH || ||| || ||| OH OH OH OH OH OH OH OH OH OH
PEGylation