NEWS: Our Blog

IN VITRO MODELS FOR PREDICTING TRANSPORTER MEDIATED DDIs

Aug 26, 2022 8:46:13 AM / by Alliance Pharma posted in Philadelphia, DDI Assessment, Drug Transporter, DMPK

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IN VITRO MODELS FOR PREDICTING TRANSPORTER MEDIATED DDIs
Membrane transporters play a critical role in drug-drug interactions (DDIs). Both major types of transporters — efflux and uptake — can affect absorption and drug disposition in the body. Transporters can impact drug safety and efficacy, acting alone or in concert with drug-metabolizing enzymes.

The picture of what is happening in the body gets increasingly murky when patients take more than one drug, which is increasingly common. To obtain insights of how DDIs occur and what their consequences might be, it is vital to conduct in vitro studies and generate the data required to determine whether clinical DDI studies are needed. This is an area where Alliance excels.

In Vitro Assessment of a Transporters’ Potential Role in DDIs

The full picture of a new drug’s DDI potential involves to determine:

  • How a new drug is absorbed and eliminated
  • How enzymes and transporters contribute to its disposition
  • How to characterize the drug’s effects on enzymes and transporters

Properly designed in vitro transporter studies can support a prediction of whether a DDI through transporters is likely to be clinically relevant, and Alliance has all the tools needed to thoroughly evaluate potential transporter mediated DDIs. We use Caco-2 cells expressing efflux transporters such as P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), HEK293 cells transfected with uptake or efflux transporters including OATP1B1, OATP1B3, OAT1, OAT3, OCT1, OCT2, MATE1 and MATE2k, MDR1-MDCKII cells, and HEK293 membrane vesicles expressing P-gp, BCRP and BSEP to study transporters, which are outlined in FDA and EMA guidelines.

Helping Clients Solve Problems and Make Decisions

Our clients’ primary need regarding drug transporters is to determine whether their drug candidate is a substrate or inhibitor of a transporter, and if that is clinically relevant. Clients need to know (1) when to do these experiments and (2) how comprehensive the experiments need to be.

(1) In a clinical program, it is crucial to know when to address questions about transporter mediated drug-drug interactions. That decision is driven by the clinical study design and the timing of the in vitro evaluation — which itself can vary based on the therapeutic indications of the investigational drug. For example, if the intended population is likely to use statins, the sponsor should examine their investigational drug’s potential to interact with OATP1B1/1B3 before initiation of clinical studies in patients. Without a question, the co-medications patients are taking factor heavily, so companies must decide how restrictive they want their studies to be. To avoid these transporter studies early on (Phase I), companies can recruit and enroll only healthy volunteers with no current co-meds prescribed to them. However, when a drug is to be administered to the target patient population — patients actually affected by the disease the drug is being developed to treat — it is critical to consider issues such as co-meds, potential renal or other impairments, and the transporters of which the new drug is a substrate and/or inhibitor.

(2) The comprehensiveness of a transporter study is influenced by the relationship between predicted in vivo drug concentrations and the concentrations at which the drug is interacting with the transporter in vitro. For example, if you have a drug that is an inhibitor at a very high concentration, but that concentration will never be reached in the clinic, you might be able to justify forgoing additional testing. (FDA decision trees for this and related questions are available here.) In vivo protein binding and the free fraction of drug in the in vitro system are important data points also to understand and characterize. These are the kinds of decisions Alliance helps our clients make.

The Key Factors in Choosing a Suitable In Vitro Test System

The appropriate model for in vitro testing depends on the desired output. For example, if you need a rapid turnaround but don’t require detailed characterization of a transporter interaction, rapid screening assays are sufficient. For more comprehensive characterization to determine whether to conduct a DDI study, HEK 293 singly transfected cells or the Caco-2 cell line are very useful for characterizing the kinetics of an interaction in detail and can be used to get the output needed. Choosing the right model to deliver the most accurate prediction of how a new drug will behave in vivo is a critical step.

Validating a Model and Experimental Conditions

Know inhibtiors and substrates are used to validate the transporter study models and experimental conditions and characterize assay performance. In other words, the most well-characterized substrates in the literature — substrates that have been studied in vitro and in vivo — are used to establish the assay initially, and then the assay quality is consistently evaluated to determine that the assays are running properly. This approach allows us to build up a history of in house data and continuously assess model performance.

How Alliance Stands Out in This Area

Alliance’s range of models from cell lines to singly transfected systems provides diverse options for our clients and provides rapid turnaround or comprehensive kinetic characterization. With these models and expertise, Alliance Drug Metabolism Services is able to help our clients answer their most challenging and important questions. The bioanalytical support from the strong Bioanalytical teams at Alliance ensure the study data integrity and dependable timelines. Very few of our competitors have this diversity of models up and running routinely, setting us apart.

One unique advantage for Alliance is our in-house expertise using Caco-2 cells. This is a polarized cell line expressing efflux transporters such as P-gp, BCRP and MRP2 that is physiologically similar to human small intestine cells, making it relevant to in vivo studies. The Caco-2 cell model is a valuable tool for evaluating P-gp and BCRP-mediated transport and assessing potential DDIs.

When it comes to DDIs, drug transporters continues to evolve, and new transporters are frequently identified. More and more drug transporters have been found to be clinically relevant. Alliance is always ready to adopt and implement new models when transporters are identified as physiologically relevant to human. We also have the expertise to characterize new transporter study models lacking of regulatory guidance, for example, our recent adoption of a bile salt export pump (BSEP) model, which does not yet appear in FDA guidance.

Alliance’s team members have a unique and deep understanding on how transporter-mediated DDI studies should be designed and executed. We understand how transporters behave, how they interact, and how they can cause clinically relevant DDIs. Whether a new potential drug is at the early discovery stage or the late developmental stage, Alliance can help our clients in the transporter studies. Visit the Alliance website to explore our capabilities and expertise.

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Ampersand Invests in Alliance Pharma, Leading Bioanalytical CRO

Nov 19, 2021 10:23:36 AM / by Ryan Klein posted in Leadership, Feng Li, CEO, success, Growth

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WELLESLEY, MA, November 18, 2021 /PRNewswire/ -- Ampersand Capital Partners, a private equity firm specializing in growth equity investments in the healthcare sector, has announced an investment in Alliance Pharma (“Alliance”), a global leader in large and small molecule bioanalytical services. Headquartered in Malvern, PA., Alliance provides a full suite of discovery bioanalytical, DMPK, regulated bioanalysis, biomarker, LC-MS/MS, immunoassay, cell and gene therapy, and protein characterization assays. These capabilities support preclinical-through-Phase 4 studies run by a global customer base comprised of leading pharma and biotech companies.

Dave Patteson, Partner at Ampersand, stated "We are very excited to complete this investment in Alliance and partner with Founder and President Frank Li, who will remain a significant shareholder in the Company. Ampersand’s goal is to help Alliance execute an aggressive growth strategy that will expand the Company’s global reach, scientific capabilities, and operational capacity.”

Frank Li, President of Alliance, stated "We are delighted to have Ampersand invest in Alliance. Ampersand’s deep industry expertise, broad network, and capital resources will fortify Alliance Pharma’s position as a market-leading global specialty CRO."


About Alliance Pharma

Founded in 2008, Alliance is a contract research organization (CRO) that specializes in advanced bioanalytical research services for both small and large molecule drugs, as well as drug metabolism studies to support pharmaceutical and biotechnology companies’ drug discovery and development programs. Alliance Pharma provides: quantitative LC-MS/MS analysis of small molecule drugs, metabolites, biomarkers, protein, peptides and oligonucleotides, as well as protein characterization services; immunoassay of proteins and antibody drug conjugates;immunogenicity assays (anti-drug antibody screening, confirmation, titer assessment, and Nab determination): cell-based bioassays; in vitro and in vivo drug metabolism and pharmacokinetic studies.

Alliance’s mission is to build a trusted partnership with our partners & clients to support their successful drug development programs. Alliance’s business philosophy is based on a foundation of trust, professional ethics, scientific excellence and regulatory compliance.


About Ampersand Capital Partners

Founded in 1988, Ampersand is a middle market private equity firm with more than $2 billion of assets under management dedicated to growth-oriented investments in the healthcare sector. With offices in Boston and Amsterdam, Ampersand leverages its unique blend of private equity and operating experience to build value and drive superior long-term performance alongside its portfolio company management teams. Ampersand has helped build numerous market-leading companies across each of the firm's core healthcare sectors. Additional information about Ampersand is available at ampersandcapital.com.

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> Our Expansions and Added Resources in 2020

Nov 16, 2020 9:00:00 AM / by Ryan Klein posted in Pharmaceuticals, Conferences, DMPK

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How has 2020 treated your company? It has been an odd year to say the least, and we are extremely grateful for the opportunities we have had in our field to work alongside and support companies, such as yours, providing medications to help people live the fullest life possible.

While we have been working with our clients on their exciting projects, we have also been working internally on growing! During the past 12 months we have expanded our footprint and capabilities within our current facility as we continue to build trusted partnerships with you – our clients! We have done this by adding the following:

Space

  • 3,000 square feet of additional office space
  • Three additional laboratories that cover approximately 1,600 square feet
  • OHC 3 – 5 lab facility
  • A BSL3 lab facility will be completed in the fourth quarter of 2020

Equipment

  • Two Agilent 6495 QqQ LC-MS/MS instruments
  • Two Agilent 8900 Triple Quad ICP-MS instruments
  • Validated Agilent Mass Hunter software
  • Two Sciex 6500+ LC-MS/MS instruments
  • New Gyrolab xPlore
  • Upgraded Watson LIMS to version 7.6.1
  • One Agilent Bio-Inert UHPLC

Staff

We have also seen a 15% growth in our staff! We continually look for scientists, lab assistants and techs, IT specialists, software developers, business development/marketing, quality assurance, fellows, and report reviewers for example. We invite you to review our openings on our website and consider joining our role serving the pharmaceutical, biotech, and agrochemical industries, as well as academia.

We look forward to working with you in the months wrapping up 2020 as well as in 2021 with our increased space, resources and staff! Please reach out to us at 610-296-3152 or info@alliancepharmaco.com to discuss how our space, equipment and staff can best meet your drug development and drug metabolism project needs.



Any further questions or need an answer?  Please contact Ryan Klein at rklein@alliancepharmaco.com or 919-801-3146.

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> Making the Most of Your Virtual Conference Experience

Nov 10, 2020 2:00:00 PM / by Ryan Klein posted in Pharmaceuticals, Conferences, DMPK

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Remember when we used to meet in person for our conference experiences? We would gather at conference centers and hotel complexes to attend great seminars, to have insightful conversation, to reconnect with old friends and to make new friends. Everything has changed this year as we all continue to adapt to the COVID-19 pandemic. We understand that many of us are sad about what we may miss, but we are thankful for the technology available that allows us to still attend conferences virtually. We know great interactions, education and conversations can still occur even if it has to happen online!

Our latest blog offers tips on how you can make the most of your virtual conference experience. We hope these tips help you as you adapt to our “new normal”. We look forward to seeing and meeting with you during virtual events in the months ahead. Thank you for the trust you put into us with your drug development and drug metabolism projects!

Tips for attending a virtual conference

We are all used to packing our bags, traveling and preparing for our interactions when it comes to attending conferences. COVID-19, however; has changed all of that this year. As many conferences are turning into virtual events, we thought we’d highlight some ways you can get the most out of your virtual conference experience.

  • Prepare mentally for attending the conference. You don't want to just "show up" online. Be prepared with any prep work you would have done prior to an in-person conference. This includes being aware of any potential distractions surrounding you, including your email and phone notifications. It’s tempting to try and multi-task in this circumstance, but do what you can to stay focused on the meeting, just like you would if you attended in person.

  • Be bold. Just as you wouldn't attend an in-person conference and stay in your room the entire time, except for a session or two, you must also be bold in these virtual settings. That means being willing to ask the questions, make the introductions and network. Many virtual conference platforms are offering video chats. Don’t be afraid to ask someone for a face-to-face call.

  • Be active on social media. Social media is a great tool for staying connected and asking questions before, during and after a virtual conference. If you aren't comfortable asking in a live event, use that hashtag to ask your questions on social media channels. Not sure you want your question in public spaces? Follow the social media channels of the conference host, speakers, panelists, sponsors, vendors and attendees, and use those direct message features or find the contact info for that direct email or phone call.

  • Take time to stroll. At a conference, you may pick up a cup of coffee and stroll through the booth areas. Do the same with virtual booth access. Vendors are supporting the virtual meeting through funding and their presence at the meeting. Provide some support for them by visiting the virtual booth and learning about their services in between presentations and during the times dedicated to the virtual exhibit hall.

  • Be patient! Just as you are learning to now navigate a virtual conference, so are all the other attendees, vendors and hosts. There will be times when technology doesn't work right, people forget to mute themselves or even random hiccups occur. Some of it may even feel awkward. Through it all, remember we are all working to adapt in the midst of all the changes COVID-19 has brought this year.

  • Don’t leave early. That last session or final connection may just be the one that makes a difference for you and your company. Make sure to take full advantage of the entire schedule and resist the temptation to head to the couch early.

You can find our conference schedule on our website and our social media channels. Please remember we may not have a table set up around the corner of the conference hall, but we are here to answer any of your questions and welcome your conversation. Visit us at our virtual booth and chat with our team through the virtual conference links at our scheduled events. You are also always welcome to contact us at 610-296-3152 or info@alliancepharmaco.com.

We look forward to seeing you at that next virtual conference!


If you’re going to any upcoming conferences and would like to meet with us, please contact Ryan Klein at rklein@alliancepharmaco.com or 919-801-3146.

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> POSTERS: 12th International ISSX Meeting

Nov 15, 2019 10:30:00 AM / by Ryan Klein posted in Pharmaceuticals, Conferences, DMPK

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Seven hundred scientists, students, and exhibitors convened at the Oregon Convention Center in Portland, Oregon, USA this past July 28–31 for ISSX2019.

Alliance Pharma is proud to have presented the three posters below.
Click poster images below to download a PDF file.

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If you’re going to any upcoming conferences and would like to meet with us, please contact Ryan Klein at rklein@alliancepharmaco.com or 919-801-3146.

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> University of Wisconsin Land O’Lakes DMPK Conference

Sep 6, 2019 10:00:00 AM / by Alliance Pharma posted in Pharmaceuticals, Conferences, DMPK

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University of Wisconsin Land O’Lakes DMPK ConferenceThe Land O’Lakes conference series is one of the longest running and most revered teaching conferences in the pharmaceutical industry. Over the last two decades, the Land O’Lakes conferences have been developed by the division of pharmacy professional development within the school of pharmacy at the University of Wisconsin-Madison and have an impeccable reputation within the pharmaceutical industry for shedding light on cutting edge topics, bringing in knowledgeable experts and having an intimate setting which lends itself to some fantastic networking opportunities. 

We at Alliance Pharma are pleased to announce that we will be attending the 22nd annual Land O’Lakes conference on Drug Metabolism and Applied Pharmacokinetics on September 9th thru September 12th in Madison, Wisconsin. We are excited and eager to hear from industry leaders on the new advances in drug delivery, action and metabolism. This years’ meeting is titled ‘Found in Translation: Adaptive DMPK Strategies’, and will include topics focused on target mediated drug disposition (TMDD), endogenous biomarkers and more.


Target mediated drug disposition requires that we think outside the box in terms of drug distribution and elimination from the body. TMDD often exhibits nonlinear pharmacokinetics and pharmacodynamics due to the drugs unusually high affinity to a binding site.


Endogenous biomarker research, particularly in the drug transporter area, continues to rapidly evolve. Our biomarker team at Alliance is a highly experienced team with significant expertise in biomarker method development and analysis, yet we are always eager to learn more!


If you’re going to the conference and would like to meet with us, please contact Ryan Klein at rklein@alliancepharmaco.com or 919-801-3146.

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> Comparing Methods for Assessing In-Vitro Protein Binding

May 6, 2019 4:00:00 PM / by Taylor MacArthur and Jessica Lipponen posted in Pharmaceuticals

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           The extent of binding to proteins is an important parameter to assess when screening drug candidates during in-vitro ADME studies. Assessing the free fraction of drug available in the biological system is critical to understanding its distribution potential, as it is generally believed that the free fraction drives therapeutic outcomes since it is the free drug (that which is not bound to proteins) that can move to and interact with the sites of action, and high binding to proteins can affect the metabolism and elimination of the drug.

          There are three methods most commonly used to assess protein binding, and it is important to assess their advantages and disadvantages when selecting methods to use for screening studies.

Rapid Equilibrium Dialysis (RED) makes use of a device with two chambers separated by a semipermeable membrane. Spiked matrix is added and over an incubation period, the free drug comes to equilibrium across the membrane while proteins and protein-bound drug cannot move across the membrane; free fraction is calculated by comparing the concentration of drug from the protein and protein-bound drug side to the protein-free, free drug side.

#1#2

Advantages: Most common method (easy cross-comparisons), simple apparatus/setup and high throughput

Disadvantages: Requires long-term stability in the matrix (typical incubation period is 4 hours), nonspecific binding to the device/membrane, volume shifts and protein leakage over the incubation period.

Ultracentrifugation (UC) uses high-speed centrifugation to precipitate proteins and protein-bound drug from a spiked matrix; free fraction is calculated by comparing the concentration of drug in the protein-free supernatant and the intact spiked matrix.#4Advantages: Avoids membrane-related issues (nonspecific binding, leakage across membrane, etc.)

Disadvantages: Usually low-throughput, harder to control pH, temperature (conditions that can significantly affect protein binding), requires longer-term stability in matrix (typical centrifugation time is two hours), the supernatant may not be completely protein-free, leading to an artificially high free fraction observed.

Ultrafiltration (UF) uses a two-chambered device separated by a semipermeable filter. Spiked matrix is loaded into the device and centrifuged at moderate speed, during which time free drug passes through the filter while proteins and protein-bound drug cannot; free fraction is calculated by comparing the concentration of drug in the protein-free filtrate and the intact spiked matrix.

#5Advantages: Suitable for unstable compounds (typical centrifugation time is 30 minutes), quick and technically simple, high throughput/useful to screen many compounds to rank free fraction

Disadvantages: Nonspecific binding to the filter membrane or collection chamber (under-estimation of free fraction), molecular sieving may occur (liquid moves through filter more quickly than the drug, under-estimating free fraction)

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> Three Essential Features of an ELISA That Every BioPharmaceutical Scientist Needs to Understand

Apr 26, 2019 11:00:00 AM / by Nathan Fritzinger posted in Pharmaceuticals, Immunoassay, laboratory

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          An enzyme-linked immunosorbent assay (ELISA) is a chemical test to detect the presence of proteins or other molecules. The enzyme-labeled antibody binds to antigens leading to a color change that is used to quantify antigen-antibody interactions. Quantification of these concentrations requires measuring the intensity of the color change, which indicates the concentration of antigen bound to the antibody. ELISA’s include four variants: direct, indirect, sandwich, and competitive, which all have their respective advantages and disadvantages.

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Benefits of an ELISA

High Sensitivity: The concentration of both antibody and antigen can be quantified on the nanogram scale. This means that scientists can use a smaller amount of enzyme, which still produces a large scale catalytic reaction.

High Specificity: Antibodies are very specific to their targets due to their spatial configuration and chemical structure. They can easily recognize and bind to antigens similar to how a key unlocks a specific door.

Versatility: Indirect ELISA’s allow for multiple primary antibodies created in one species with the same labeled secondary antibody used for detection. Also, it’s easy to wash off any unbound antibodies by applying a detergent solution to the plate.

Clinically approved ELISA tests for diseases like HIV, Lyme disease, and tuberculosis are proving their efficacy and value to the Pharmaceutical industry.

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> AAPS Poster - #M1430-03-017 - Development of an LC-MS/MS Method for Quantification of Cytarabine in Rat Plasma.

Oct 26, 2018 6:49:06 PM / by Karen Pryor

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AAPS Poster - #M1430-03-017 - Development of an LC-MS/MS Method for Quantification of Cytarabine in Rat Plasma - ePoster presentation track: BioanalyticsForum3 - Screen 17 - Date/Time 2018-11-05 2:30

Alliance Pharma is gearing up for the #AAPS PharmSci 360 Conference in DC.  Meet Our Scientist for Poster  #M1430-03-017 - Development of an LC-MS/MS

Method for Quantification of Cytarabine in Rat Plasma. Stop by our booth #610 or schedule a meeting with us at the conference Ryan Klein 919-801-3146 or email rklein@alliancepharmaco.com.

 

It’s a great opportunity to connect with leading #pharmaceutical #scientists from across the globe!Conference Announcement Template - D

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> Building Stronger Collaborative Partnerships with Cutting-Edge Instruments

Oct 23, 2018 4:36:17 PM / by Karen Pryor posted in Pharmaceuticals, Philadelphia, bioanalytical, laboratory, Bioanalysis, Growth, 6545XT, QTOF, AdvanceBio

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Alliance Pharma is dedicated to bringing comprehensive solutions to our collaborative partnerships.  We understand the importance of efficiency and our team of scientists strive to provide the highest analytical services for your project. 

Our recent purchase of multiple Agilent 6545XT AdvanceBio QTOFs – which is specifically designed with biopharmaceutical characterization in mind - provides high resolution and sensitivity for qualitative and quantitative analyses of biomolecules ranging from peptides to intact monoclonal antibodies. The iterative peptide mapping workflow allows us to dig deep into samples with a wide range of analyte concentrations (i.e. host cell proteins and sequence variants). The Agilent 6545XT AdvanceBio QTOF maximizes our uptime by performing thousands of protein injections without degradation of performance with mass accuracies within 10 ppm for glycosylated intact proteins. 

At Alliance Pharma we believe keeping up with the latest technology helps us to provide the best services for our clients.  The 1290 Infinity II LC system when coupled with the 6545XT, represents the next generation LC for ultrahigh-performance liquid chromatography with superior reliability, offering efficient and reproducible separations of biomolecules for detection by MS or integrated UV and FLD modules. 

By using instruments that complement our scientist’s knowledge and experience we meet analytical objectives, increase productivity, and are cost-effective.  Contact us to see how our recent instrument purchase will help keep your project on track.  Call or email Ryan at 919-801-3146 or  rklein@alliancepharmaco.com for more information.

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