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Unlike the development of most small or large molecules, which typically measures only one part or the metabolite for pharmacokinetic analysis, multiple parts of ADCs need to be measured to characterize their PK properties. Therefore, a thorough understanding of the clinical pharmacology of ADCs is critical for selecting a safe and effective dose for patients. Biological Analysis of ADC In order to characterize the PK characteristics of different ADC components, several analytical methods are required: (1) ELISA determines the kinetic curves of the conjugate and total antibody. (2) TFC-MS/MS can quantify the free drugs or metabolites. (3) High-resolution mass spectrometry for in vivo drug antibody ratio (DAR) analysis. In addition, two types of ELISA immunoassays are used to quantitatively measure the ADC analytes: The first type measures total antibodies, namely ADCs with DAR greater than or equal to zero. The second assay measures drug-binding antibodies, defined as ADCs with DAR greater than or equal to 1. Other analytical methods include size exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC). Cytotoxic Payload ADC cytotoxic payloads should have the following characteristics: (1) Proper lipid solubility. (2) Stable in blood. (3) The target of payloads locates inside the cells. (4) The payload molecules are small in size, lack immunogenicity, and are soluble in water buffers so that they can be easily coupled. Currently, the commonly used cytotoxic effector molecules are microtubule inhibitors (e.g. auristatins, maytansinoids), DNA damaging agents (e.g. calicheamicin, duocarmycins, anthracyclines, pyrrolobenzodiazepine dimers) and DNA transcription inhibitors (amatoxin and quinoline alkaloid (SN-38)). Drug Antibody Ratio (DAR) Studies have shown that ADCs with a high DAR (7 to 14) are cleared faster and have reduced in vivo efficacy compared to those with a DAR lower than 6. DAR values and their effect on the stability and PK also depend on coupling locations and linker sizes. Lysine or cysteine is often modified to produce ADC. Lysine is one of the most commonly used amino acid residues to link a substrate to an antibody. Lysine is usually present on the surface of an antibody and, therefore, easily conjugated. Mylotargs, Kadcylas, and Besponsas all use lysine bio-binding technologies. And other amino acids such as cysteine and tyrosine can also be modified. For instance, ADCs such as Adcetriss, Polivys, Padcevs, Enhertus, Trodelvys, and Blenreps have been synthesized by modifying cysteine with maleimide. Linker The latest generation of linkers is more stable in the systemic circulation, such as peptide and glucuronic acid linkers. The two most common linkers are introduced below: (1) Cleavable linkers are sensitive to the intracellular environment and release free effector molecules and antibodies through catabolism and dissociation. They are usually stable in the blood but are rapidly cleaved to release effector molecules in a low pH and protease-rich lysosome environment. In addition, if the payloads are transmembrane, they can eliminate tumors through the potential bystander effect. (2) Non-cleavable linker is a new generation linker with better plasma stability compared to cleavable linkers. Since non-cleavable linkers can provide greater stability and tolerance, they can reduce off-target toxicity and also provide a larger therapeutic promise. Immunogenicity In 11 clinical trials of eight ADCs, the baseline incidence of anti-drug antibody (ADAs) ranged from 1.4% to 8.1%, and the post-baseline incidence of ADAs ranged from 0-35.8%, values within the range of therapeutic monoclonal antibodies. In general, the incidence of ADA of ADCs is less in patients targeting hematologic cancer than in patients targeting solid cancer; Most ADA is specific to the monoclonal antibody domain of the ADC. Furthermore, in most patients, the hapten-like structure of these ADCs does not generate any more risk of an immune response than therapeutic monoclonal antibodies. ADC Pharmacokinetic Model The application of models can integrate PK, efficacy, and safety data to meet the needs of ADC drug development at different stages, such as target selection, antibody affinity, linker stability, animal-to-human extrapolation, dose selection and adjustment, exposure-response relationships, DDI studies, etc. ADC's kinetic model is complicated due to its multiple clearance pathways (dissociation and catabolism) and complex PK characteristics of various analytes. Future Vision Clinical pharmacology plays a very important role in ADC drug research. An in-depth and comprehensive elucidation of the PK/PD characteristics is crucial to promote the development of ADC drugs with lower toxicity and higher efficiency. Backed by advanced biological analysis techniques, researchers and clinicians will witness wider applications of ADC drugs in the field of cancer treatment sooner or later.

Cherwell has published an update to its guide on “Environmental Monitoring Processes and Validation”, incorporating specific detail on the new version of EU GMP Annex 1. This aims to help sterile medicinal product manufacturers with reviewing and improving their Environmental Monitoring (EM) programs in preparation for compliance with the revised guidelines by August 2023. Available to download from Cherwell’s website, the new eBook follows Cherwell EM experts’ detailed review of all changes included in the extensive 59-page 2022 revision of these key regulations governing the manufacture of sterile medicinal products in the UK and Europe. Cherwell’s updated guide adds new and more detail on a number of areas including: Risk-based facility management; Zero CFU; Continuous EM; Trends monitoring; Personnel training; and VHP fumigation. Cherwell’s EM eBook also includes an update to its section discussing how to achieve Annex 1 compliance in EM processes and programs by taking a practical and bespoke approach to understanding the level of compliance required by individual facilities. This can be achieved by breaking down preparation into four steps, starting with a full EM audit and analysing every aspect of a facility’s current EM program. To help further understanding, the guide also offers examples of best practice EM programs within both industrial pharmaceutical and commercial scale hospitals and aseptic compounders. In order to maintain industry knowledge and stay up-to-date with the latest developments, including regulations such as the new EU GMP Annex 1, in addition to in-depth scientific reading, Cherwell’s team regularly attends industry conferences and events. This enables the team to work in partnership with and advise customers when reviewing their EM and contamination control strategies. “At Cherwell we strive to best support our customers in making the right decisions associated with combining methods and instruments to cost effectively achieve the best results for their individual EM and validation protocols,” said Thomas Parkhill, Microbiology Sales Specialist, Cherwell. “In this eBook, we offer a consultative guide to the most business-efficient EM measures organisations can adopt to comply with GMP Annex 1, and practical steps they can take to create the ideal EM process for their specific business needs.” Hamish Hogg, Microbiology Product Specialist, Cherwell, added, “The sheer volume of new content in the much-revised GMP Annex 1 alone does present a challenge for businesses wanting to comply; they might be overwhelmed by the choices needed in order to determine the right path for them. That’s why we aim to work in an advisory role with our customers and assist them by drawing on the extensive knowledge and expertise held within our team. Our newly updated EM guide is just one such example.” With over 50 years of experience, Cherwell has built a reputation within the pharmaceutical and healthcare sciences industry as a provider of high-quality products to meet the specific requirements of environmental monitoring and process validation. In addition to the ImpactAir continuous viable air monitoring solutions and SAS microbial air samplers for active environmental monitoring, Cherwell manufactures and supplies the Redipor® prepared media range, comprising agar plates, bottled media, vials, broth bags and ampoules. Cherwell’s updated eBook on “Environmental Monitoring Processes and Validation” detailing the latest Annex 1 revisions, is available for download at: NEW Environmental Monitoring Processes & Validation Guide.

For monitoring of thermal effects, physical and chemical reactions and much more directly in the grinding bowl! With the new EASY GTM Gas Pressure and Temperature Measuring System you can turn the Planetary Mills PULVERISETTE 5 and PULVERISETTE 6 of the FRITSCH classic line into analytical measurement systems. Due to the continuous measurement of gas pressure and temperature, thermal effects as well as physical and chemical reactions or pressure variations can be monitored directly in the grinding bowl. For this, simply insert the grinding bowl with the radio transmitter integrated in the lid, without any modification to the mill itself. The monitored data is transmitted to a computer via a receiver. The EASY GTMControl software included in the scope of delivery generates a graphical presentation of the measured values, which can also be offered as a PDF report and transferred to an Excel™ table. The EASY GTM-System provides valuable information - Investigations in the area of mechanical alloying for the production of new amorphous and nano-crystalline materials - Monitoring and optimisation of grinding processes in industrial applications The measurement of the grinding bowl temperature allows an integral statement on the process variable temperature, which takes account of the effects of all friction, impact and transformation processes. The continuous and highly sensitive measurement of the gas pressure in the grinding bowl enables the detection of very rapid reactions. The measured gas pressure describes, among other things, the interactions of the gas with the surfaces created during grinding (adsorption and desorption of gases). Extremely rapid phase formations can be observed for the first time IN SITU as an adiabatic process without heat exchange with the system. TECHNICAL DATA - Temperature measurement range of the transmitter component: –40 to 110 °C - Resolution of pressure signal: 1 mbar - Permissible pressure in the grinding bowl: 0 – 20 bar – measuring range up to 30 bar - Operating time with fully charged battery: approx. ½ year continuous operation (depending on operating temperature) - Radio transmission: 2.4 GHz standard

Lipase (glyceride hydrolase) belongs to the class of carboxyl ester hydrolase, which can gradually hydrolyze triglycerides into glycerol and fatty acids. Lipase exists in fat-containing animal, plant and microbial (such as mold, bacteria, etc.) tissues. Including phosphatase, sterolase and carboxylesterase. Fatty acids are widely used in food, medicine, leather, daily chemicals, etc. Lipases are widely present in animals, plants, and microorganisms. Plants containing more lipase are seeds of oil crops, such as castor bean and rapeseed. When oil seeds germinate, lipase can cooperate with other enzymes to catalyze the decomposition of oils and fats to produce sugars, and provide nutrients and energy necessary for seed rooting and germination. The pancreas and adipose tissue of higher animals contain more lipases in animals. The intestinal juice contains a small amount of lipase, which is used to supplement the insufficiency of pancreatic lipase for fat digestion, and the gastric juice of carnivorous animals contains a small amount of butyric acid glyceryl esterase. Properties of lipase Lipase is a class of enzymes with a variety of catalytic abilities, which can catalyze the hydrolysis, alcoholysis, esterification, transesterification and reverse synthesis of esters of triacylglycerides and other water-insoluble esters. In addition, it also shows the activities of other enzymes, such as phospholipase, lysophospholipase, cholesterol esterase, acyl peptide hydrolase activity, etc. The different activities of lipase depend on the characteristics of the reaction system, such as promoting ester hydrolysis at the oil-water interface, and enzymatic synthesis and transesterification in the organic phase. The properties of lipase mainly include the optimum temperature and pH, temperature and pH stability, and substrate specificity. So far, many microbial lipases have been separated and purified, and their properties have been studied. They are different in molecular weight, optimum pH, optimum temperature, pH and thermal stability, isoelectric point and other biochemical properties. In general, microbial lipases have a broader pH, temperature range, high stability and activity than animal and plant lipases, and are specific to substrates. Lipase is one of the important types of industrial enzyme preparations, which can catalyze reactions such as lipolysis, transesterification, and ester synthesis. It is widely used in oil processing, food, medicine, daily chemical and other industries. Lipases from different sources have different catalytic characteristics and catalytic activities. Among them, the large-scale production of lipases with transesterification or esterification functions for organic phase synthesis is of great significance for the enzyme-catalyzed synthesis of fine chemicals and chiral compounds. The main use of lipase Lipase from microbial sources can be used to enhance the flavor of cheese products. The limited hydrolysis of fat in milk can be used in the production of chocolate milk. Lipase can make food form a special milk flavor. Lipase can prevent the taste of baked goods by releasing monoglycerides and diglycerides. The degreasing of bones in the production of gelatin needs to be carried out under mild conditions, and the hydrolysis catalyzed by lipase can accelerate the degreasing process.

Probiotics are a type of active microorganisms that play a role by improving the balance of the host's intestinal microflora. They can produce definite health effects to improve the host's microecological balance and play a beneficial role. In 2001, the World Food and Agriculture Organization (FAO) and the World Health Organization (WHO) also defined probiotics as follows: Live bacteria that can play an effective role in the health of consumers by ingesting an appropriate amount. Bacteria or fungi in humans and animals mainly include: Clostridium butyricum, Lactobacillus, Bifidobacterium, Lactobacillus acidophilus, Actinomycetes, Yeasts, etc. The intestinal flora is the largest pool of human bacteria and the largest micro-ecosystem. There are about 1,000 kinds of bacteria in the intestine, 100 trillion bacteria, 10 times the total number of human cells, including bacteria, fungi, archaea, and native animals and plants (even viruses) and many other types of microorganisms. The intestinal flora is inseparable from the human body and is a unity of symbiotic and mutually beneficial relationships, and is also called the “second genome” of humans. The total weight of bacteria in the intestines can reach 1-1.5 kg, and more than 50% of the weight of feces excreted by the human body every day is composed of bacteria. In the past two decades, due to the improvement of sequencing technology, scientists have paid more and more attention to the research of intestinal flora. It is found that probiotics can help digestion and absorption, promote the balance of intestinal flora, and maintain human health. A lot of evidence shows that different people have different composition of intestinal flora. Diet, drugs, and environmental factors can all affect the composition of an individual's intestinal flora. The health effects of probiotics on the human body are not limited to the intestines, but also have broader effects, such as immune balance regulation, and also affect organs such as the brain and liver. Disturbance of intestinal flora is related to many chronic diseases such as diabetes, obesity, depression and so on. In recent years, scientists have conducted a lot of research on the usefulness of intestinal flora to humans, revealing how intestinal microbes can help improve the host's body health and help prevent diseases. Researchers have discovered that Lactobacillus rhamnosus GG (LGG), an important member of probiotics, can help treat intestinal problems and respiratory infections. They also found that the bacteria can help lose weight. So can LGG really bring benefits to mankind? Researchers from the University of Maryland School of Medicine published the latest research results in the journal mBio and found that LGG can play a role as a promoter to modify the activity of other intestinal bacteria. The study tested 12 subjects. Participants took LGG twice a day for 28 consecutive days to analyze the intestinal flora before and after the treatment. It was found that the intake of LGG can increase the expression of many genes, thereby promoting the growth of other intestinal flora, including Bacteroides, Bifidobacterium and other flora. These flora are considered to be beneficial to humans, including promoting the healthy development of the immune system. Dr. Rinki Murphy of the University of Auckland found that a very low-calorie diet and intake of probiotics may help effectively inhibit the occurrence of type 2 diabetes. The effect of specific probiotic strains in inhibiting diabetes can help effectively reduce the occurrence of gestational diabetes, and can improve the patient's insulin sensitivity and body weight. Inhibiting type 2 diabetes, intermittent fasting can magnify the beneficial effects by supplementing Lactobacillus rhamnosus probiotics. Regarding the role of supplementing probiotics, there have been thousands of related studies published in the past few decades, and the number of clinical studies related to probiotics has also been increasing year by year. There are many positive experimental results for intestinal regulation, immune regulation, alleviation of allergies, cancer suppression, and women's health.