Mr. Brian Hilton: Hello, and welcome to Almac's presentation on blinding techniques and analytical support for clinical trials. The goals of today's webinar are to gain insight on how blinding is an integral part of the clinical trials to remove both patient and investigator bias as well as gain a better understanding on blinding techniques associated with analytical support services for drug product

I am Brian Hilton, Production Manager of Almac Clinical Services, and the analytical portion will be presented by Aeri Park, Director of U.S. Operations for Almac Analytical Sciences

Welcome. My name is Brian Hilton, Production Manager with Almac Clinical Services. Within this segment, we will evaluate the blinding considerations during the over encapsulation processes

Blinding, as found in the roles and guidance for pharmaceutical manufacturers and distributors Annex 13 is a procedure in which one or more parties to the trial are kept unaware of the treatment assignment. As it relates to an investigational medicinal product, blinding shall mean the deliberate disguising of the product in accordance with the instructions of the sponsor

Without maintaining a proper blind, patients who know what treatment they are receiving might harbor favorable expectations or increased apprehension, thus skewing the study data

Over encapsulation is a process by which one or more products are assembled into a hard gelatin shell, which may or may not be filled with an inactive bulk agent or excipient

The ability to maintain blindness throughout the complete clinical trial processes must not only focus on the visual aspects of the product but also take into consideration the other human senses. Taste and smell are by far the two most difficult senses to blind against

Many years ago, I learned of a study in which the client's product had a strong odor. In order to mask the smell, the client inserted a single table into a canister desiccant for use during assembly of bottles. The bottled product inherited the pungent smell, thus maintaining proper blindness

Unfortunately, not all products have a blinding solution

Within clinical trials, over encapsulated material will undergo further processing. Based on the client's protocol, this will often be assembly into foil blister packs or bottling operations

Just as with many companies across the globe experiencing economical struggles, the pharmaceutical industry is feeling the pressures to be more effective, efficient and leaner. Whether it be the rising cost of R&D, the astronomical funding required to bring a new product to market, the potential need to purchase specific equipment and tooling for use during your clinical packaging, employing healthcare costs, large pharma to small bio tech companies are being forced into outsourcing work

Are the chosen materials suitable for the target patient? To elaborate, is the size capsule suitable for children, seniors

Manufacturing and packaging of clinical supplies only continues to increase in complexity. Over encapsulation, bottling, blistering and/or labeling each possesses its own set of challenges. It is not simply a case of variation of batch size, multiple setups with the required process checks, but variation in capsule size, flow of excipient used and the shape or dimensions of the input product that pose further challenges

Complexity of dosages may require specific unique solutions. Certain strengths in the comparative product may not be available, requiring insertion of multiple units of the same or differing product

Purchasing of a global comparator should focus on how readily available the product is--additionally, what quantities will be required as the clinical trial progresses

Are expiry dates lengthy enough to allow for packaging, testing, material release and patient enrollment

The selection of backfill should pose no impact to the over encapsulated product and should allow for efficient processing

We will further evaluate both the comparator and backfill selection in upcoming slides

Over encapsulation is the most widely accepted blinding technique. Why? Capsule shell suppliers offer a vast array of opaque colors that can be used to obscure input components

Your final assembled unit is visually identical from one product to the next, thus reducing influence from the trial participants, investigators, assessors

The over encapsulation process allows for one or more products to be placed into a capsule shell. This aids in configurating [sp] dosage that may or may not be available in the market

Processes and testing must show that product quality has not been altered and requires justification of expiry date as each component, the input unit, shell and backfill each retire expiry date

The analytical segment of this presentation will assist in guidance to correct expiry dating

Patient compliance requires maintaining a blind that resists tampering and clearly reflects when tampering has occurred

Rapid unblinding to identify the product in case of an emergency

More often, the comparator selection will seek to take on a market leader. Let us not think that this selection does not pose its own challenges

As discussed previously, consider the lead time for the delivery and expiration dates of your comparator product. Can the comparator be used across global markets

Once consumed, will the capsule shell and added backfill break down in suitable timing to allow release of the product? Is the physical shape and size of the product able to fit into suitably sized capsule shells

This again relates back to your target patient. For example, a stroke victim may have difficulties swallowing a large capsule

Is the comparator product too brittle to effectively use automated processes as this could jeopardize timelines and costing

Suppliers can produce consistent stocks of hard gelatin capsules where the range of diameters and length allow many products to be over encapsulated. It is important to ensure the color of the capsules chosen completely obscures the input material

Confirm that the comparator or the IMP [sp] fits within the capsule shell without distorting the shape. This may require splitting of the input units

Is the shell a universally accepted color? This is vitally important as some regions may not allow certain colored capsule shells to enter their borders

Provide thorough research to understand and consider what the acceptable daily intake is for both the capsules and the colorings

Within the DB series line of capsules, it offers a smaller size, which helps to improve compliance, completely overlapping of the capsule top to the capsule bottom

Once closed, the locking mechanism makes the unit difficult to open without damaging the exterior shell

Numerous sizes are available and offers a globally approved color selection, allows for high speed production capabilities

Blend and/or excipients are primarily used to prevent rattling, fill capsule void and manufacture of placebo to match capsules containing excipient only

What excipient should be used? It is important that this be inactive, often is the same excipient that is used during the formulation of the active product or the comparator product

Off the shelf products that are available include powder, pellets, sugar spheres. Blends may also be utilized. This may be a lactose with a magnesium stearate

Excipient chosen may impact upon machinery type used. Dependent on the quantity of material to be over encapsulated, shape of the product and the number of input components, there are a series of processes that may be used, including manual, semi-automated or automated processes

Manual encapsulation operations are most suited for smaller size batch runs including over encapsulation of one or more units, placebo to match capsules containing a placebo tablet or a backfill only and can accommodate uniquely shaped tablets that do not lend themselves to the automated fill process

Although manual processes allow for a quicker setup, the manufacture of capsules is a time consuming process with challenges of obtaining consistent weights across all units

Semi automated encapsulation becomes most relevant when manufacturing small to large size batch runs that include assembly of placebo to match capsules or insertion of one or more active solid dosages into capsules that may or may not contain backfill

Limited capabilities exist to automate the unit insertion depending on size and shape of the unit

Semi-automated processes allow for an increased rate of capsule assembly by use of automated fill processes or tooling assist [sp] processes. This process is also best suited when the input unit needs to be broken in half to properly fit within the capsule shell body

Average capsule weights become more consistent by use of semi-automated and fully automated equipment

Fully automated encapsulation is recommended when manufacturing medium to very large size protocols that include assembly of placebo to match capsules or insertion of an active solid dosage into capsules that may or may not contain backfill

The automated equipment allows for the most efficient rate in capsule assembly as rates may increase to nearly 42,000 capsules per hour

Certain automated equipment may allow for the insertion of multiple units of the same product into each capsule body

Average capsule weights become increasingly consistent through the addition of a controlled dose of excipient to each capsule instead of simply flood volume filling

Following the completion of over encapsulation processes, check weighing should be performed to confirm uniformity of weight and to detect any potential miscompiled capsules. Additionally, when having utilized a backfill, capsules should pass through a deduster to remove any powders from the outer capsule shell

A final check should be to pass all units through metal detection equipment

The next portion of today's presentation will focus on bottling and how to maintain blindness between treatment groups

Within this segment, we will evaluate the blinding considerations during the bottling, blistering and walleting processes

Bottling is likely the simplest, most efficient method to assist in maintaining blind during the patient dosing

As many variations and bottling components exist between suppliers, it is recommended to source efficient quantities of each component from not only the same supplier but also the same batch for each campaign of your clinical trial

Within your study, consider if bottles of various dosages will be compiled side by side within a patient kit to ensure blindness can be maintained beyond the bottling processes

Visual variations in the way products are bottled, capped, sealed or labeled may break the study blind. A 60 CC bottle may be tall and thin or short and stocky. Consider the destination in which your supplies will be distributed. A thicker bottle density will withstand higher pressures during air transits

Differences in text on the cap and/or seal liner can be a quick visual identifier of differing products

Although adding components in any aspect of the clinical packaging process provides further challenges to maintaining blind, each come with their advantages. The addition of a cotton rayon polyester filler can significantly reduce the risk of product breakage during transit

These fillers may be cut into various lengths, depending on the bottle size versus product fill level. However, it is also important to consider factors such as patient population or product stability when selecting a suitable filler

For moisture sensitive products, a canister or sachet desiccant is very effective in removing moisture in the head space of the bottle present at the time of packaging. While every gram of silicone or clay only absorbs one-third of its weight, the appropriately sized desiccant minimizes the risk to absorbing moisture from the product

Although blistering provides additional challenges to maintaining blind, is more complex in nature and arguably less efficient, blister packaging has its benefits

The general principle of blister packaging is to hermetically seal each tablet or capsule into its own cavity. Two basic types of pharmaceutical blisterings exist

Thermoform material utilizes a clear plastic providing transparency to the product within each cavity. Cold form material utilizes a foil as the essential component of both webs

Cavities are formed by cold stretching the foil and provides no transparency to the product once sealed

As a whole, blister packaging is believed to be better than conventional packaging as it maintains product integrity as well as protection, tamper evident as each dose is extracted one at a time, and reduces the risk of accidental misuse by the patient

Sourcing a new comparator product already contained with a blister pack may require deblistering and reassembly into new blister packs as differences in materials used may be difficult to mimic

Subsequently, different blistering equipment may or may not provide identical blister packs. A small sampling of the many different blistering materials and configurations are highlighted on the following slide

Although less efficient, manual blistering allows for the assembly of multiple products to be arranged in any configuration with any preformed base tray

Whether handling multiple, identical or non-identical products, clinical packaging vendors should have detailed procedures in place to minimize the risk of product crossover and/or contamination

Automated blistering can offer significant advantages through any assembly of blister packs. Depending on each package's equipment library, automated equipment has the capabilities to offer single or mixed product fills into multiple blister pack configurations

Various equipment options also allow for the addition of a print mat, which is an imprinted code applied to each blister pack for product identification and is only visible up until time of wallet assembly, pinhole detection and fully integrated vision systems

Manual feeding online requires the placement of the input unit directly into the open cavity

During mix filling, individual and locked product toppers are required during the mix product packaging

Numerous options exist for blister packaging of multiple products of varying size and shape to be assembled into a single blister pack

Maintaining product compliance, especially when mix foiling, is achieved through various verification systems. Pre-programmed vision systems are trained with the size, shape and color of the product and will detect an accidental miscompiled blister pack

Product may be issued in small sub-lots and reconciled more often at tighter reconciliation limits

Check weighing of all blister packs, both Thermoform and Cold Form, is utilized to assist in detecting an open cavity

X-ray verification in assembled cold form blister packs may detect a miscompiled blister pack without the need to conduct a full product recovery

Although simplified, walleting still poses challenges to maintaining blind during the assembly process. To maintain blind during assembly of wallets, careful consideration should be given to apply tracer markets, embossings or print on the blister pack versus its orientation into the wallet stock

Confirm that the print will not be visible at patients dose their medication through the push through lidding

To minimize risk of assembly of incorrect wallets, physical barriers should segregate operations utilizing more than one identical product. Additionally, sub-lots and more stringent reconciliation limits are recommended

As indicated in the below photos, variation of wallet stock, color and visibility of print codes each provides a risk to maintain blind

Manual walleting allows for many different input units to be assembled into a larger wallet stock that would not fit on automated equipment. Although allowing for any configuration, speed of operations are dictated by personnel

Semi-automated processes allow for increased efficiencies as the speed of operations is dictated by the equipment. As personnel man product fill stations along the equipment, the number of input units may be limited

Masking may be utilized to assist in correct product placement as well as visual verification prior to sealing of the wallet

Fully automated walleting equipment provides the most efficiency into wallet assembly. However, it comes at a tire [sp] tooling cost and significantly longer equipment changeover times. Automatic loading of blister packs into two piece wallets eliminates operator manipulation

Visual systems detect the presence of each blister and confirms correct orientation

Within this segment, we will evaluate the many challenges associated with blinding

Inhalers where physical differences are so extreme making blinding difficult to achieve, modifications of the inhalation device or using the same device for both treatments is not always feasible because the inhaler plays such a critical role in the disposal and subsequent action of the active ingredient into the airwaves

Just as there are many differences in the size and shape of solid oral dosages [sp], so too are there many variations in inhaler devices, as detailed here. It is these variations that present the pharmaceutical industry with so many challenges when needing to blind

Processing of inhalers may require the removal of labels, embossed details and/or ink markings

Additional blinding considerations focus on the need to closely mimic the canister pressure at time of actuation, the sound, the taste and the potential residue remaining around the mouth after inhalation

Blinding considerations may include delaying of an embossed print code, dismantling of the unit, including removal of all product labels to convert an active inhaler to a placebo inhaler. This may also include the breaking down and washing of inhalers under local exhaust ventilation

Accuhalers and Turbuhalers pose just as many challenges to the blinding process. During unit conversion, specific equipment and detailed processes may allow for the removal of the active product and reassembly of the unit now containing a matching placebo

Depending on your protocol design, blindness may not always need to be taken to the inhaler unit. Blindness may simply be maintained through the addition of study labels or assembly of units into sealed pouches or cartons

During unit conversion, modified blistering equipment is used to assemble matching placebo products. Each aspect of the unit conversion is monitored through in process and quality control checks to confirm seal, functionality and weight

Subsequent slides illustrate the unit conversion process

Blinding of Turbuhalers may require the overlay link of a printed component with blinding label or using an acetone to remove printed text

During the blinding process, contents of each inhaler are removed and weighed followed by a dismantling into each component part. The components go through several wash cycles using a series of methanol baths and Sonicators

To confirm the component wash removed all active residue, a sampling of the components is performed. Units may then be reassembled with placebo backfill matching the original weights

Continuing the trend, blinding of syringes, vials and patches provide significant challenges with even fewer viable solutions

Although it is feasible to disassemble a syringe to change out a portion of components from the original assembly, challenges still exist in regards to barrel style, product color and fill volumes

Similar challenges also hold true when working with vials and ampoules. Various sizes, shapes, fill contents, cap colors, embossings, vial wall densities, storage conditions each poses their set of blinding conflicts

A number of proven blinding methods would allow for the removal of the commercial label, removal of the print applied directly to the unit and/or a restriking [sp] of the print for complete obscurity. Additionally, application of a shrink wrap over the vial would allow for the fill content and color to be maxed

Variances in pouches include ink jet codes applied directly to the unit, tear notch assist, which is the opening feature of the pouch, as well as the seal area around the perimeter of the pouch

Pouches that are similar in size may be fully encased with any stud label for complete blinding of the units

Variance in tubes include tube size, any ink jet or embossed codes applied directly to the unit, the crimp thickness at the base of the tube as well as cap styles

Due to the limited options for blinding, it is recommended that you contact your packaging contractor for viable solutions

Within the labeling and compilation, we will explore the considerations needed during final packaging to maintain blindness

It would be pointless to have taken every precaution of maintaining blind with previous operations only to have variances in labels and/or general components on blinded study. Blinded study should provide consideration to variations in label stock, including the adhesives, the transparency of label if being used to urva [sp] label and existing label as well as label backing and/or color differences between the label stock

Considerations should also be provided to label profiles including the font style or size, the print boldness, the print indentation as well as any stray print marks

Ongoing challenges will often be present at any phase of the clinical packaging. It is the ability to recognize these differences and the precautions put in place to limit the chance of unblinding a clinical trial

Master samples are taken at the first receipt of any raw material type and are referenced as the blinded sample upon inspection of subsequent lots of that same material type

For blinded samples and first unit assemblies, a sample unit is created in the first operation of each campaign. The same unit will be used for direct comparison in subsequent operations

Packaging contractors may utilize design layouts and labeling templates to assist in providing consistency across all operations within a campaign

Routine inspections are performed by production and quality personnel, ensuring correctness of unit assembly, label placement, patient kit collations and blindness checks

The next series of slides will highlight Almac's equipment capabilities during your clinical packaging trials

Almac encapsulation equipment, the Excellulose [sp], Sinazi [sp], Modusee [sp], Schaffer [sp], MG2 and the Bosch GKF 702, Almac bottling equipment, Kalers [sp] 12 track bottling lines, Almac blistering equipment, pneumatic blister sealer, Klockner EAS, the Noex 623 and PharmaWorks TF3, Almac's wallet equipment Shuttle Zeds [sp], Rotary Zeds [sp], Automated Inline Walleting Press. Thank you

Ms. Aeri Park: Thank you, Brian, for your thorough presentation on the encapsulation on all the packaging options

As Brian introduced, my name is Aeri Park. I'm Director of U.S. Operations in Souderton, Pennsylvania, and I'm in charge of providing all the analytical services for your over encapsulated blinded commercial products

So, I will talk about in this segment what type of analytical testing is required for the blinded products as well as why we have to test those materials and different testing techniques and some of the problems that we encountered and how we overcame them. I also talk about stability testing and various other aspects such as timelines. So, stick with me, and we'll talk about that shortly

Hello. My name is Aeri Park. I am a Director of U.S. Operations in Almac Group in the U.S., providing all the services for your blinded material, which is over-encapsulated products, how to perform the analytical testing, why we have to perform the analytical testing and what type of testing that's required

So, I will talk about all of this today. But, first, when you talk about blinded material, obviously, that's over-encapsulated material, the key considerations we have to consider are, first of all, the true product because there are more than one types of manufacturers for the same drug products and different sizes and shapes

Because they have to go into another capsule shell, size and shape can be important, impacting the size of the final capsule shell you have to put in, and also amount of excipients you have to fill in as a backfill. And also, you have to consider the type of excipient because that may impact performance of your blinded products

The capsule shell is obviously also very important, and we will talk about a little more of this later

Now, analytical data is very important because your study results have to have the right test to make sure you have the right placebo that's labeled as placebo and the active is right active and the right amount of active if you're using multiple potencies

Now, today, we'll mostly talk about oral dosage forms. But, you know, same principle can be applied to many different variety of products, which I'll have some examples of that later during the presentation

So, there are just two pictures here. If you look at the top picture, it shows some tablets and some of the over-encapsulated products and some capsules

So, when you first blind your product, either it's tablet or capsule, putting into another big capsule and fill it with the backfill, the most important testing to be performed are listed here. It's identity, because you have to make sure the placebo is indeed just a placebo, active contains your product, potency because if you have a multiple strength of the product you are blinding, you have to make sure the potencies are correctly prepared

Now, the most important test for all blinded products is performance. Because you have--even though you have taken commercial product, the commercial product are now inside another capsule with another backfill. So, you don't know if this tablet or capsule will now perform exactly same way as intended as the original commercial product

That's why performance is important, and that is tested by something called dissolution testing. And we'll talk about that in detail later

Now, if you look at the bottom picture - so, all these over-encapsulated blinded products are put into a packaging. The picture shows example of a blister packaging. That's put into another box

So, once it's packaged, either blisters or bottles--again, identity has to be checked, make sure the right placebos and right potencies and right products is in the product, in the blister package or bottles

Now, now it comes shelf life--the original commercial product we purchased to blind may have its own shelf life. But, now we have taken it, put it into another capsule, filled it with a backfill, and then packaged it again differently. So, new shelf life has to be determined, and this is determined by something we call stability testing

And so, during the duration of clinical trials, stability testing needs to be performed to ensure these products, these blinded products are still safe and okay to use

Here, it's a very simple scheme showing how these products are tested at different time points. So, if you think about the very simplistic view, initial--very initial point, the original commercial products--let's say it's tablet, it's over-encapsulated, that’s the first blinding step

Once it's over-encapsulated, you have these bulk products of capsules. Now, this is a good point to do the testing, and I'll explain to that in a few minutes

So, these bulk products can be tested for ID. That's identification, make sure there's a right product in it, placebo's a placebo, there's only backfill in it obviously for placebos, assay and impurity--assay is for testing for potency, impurity obviously important for later. I will talk about why impurity is important

Dissolution--I talked about the performance, make sure that blinding did not alter any of the performance of the original product. And microbiology - this is to ensure that there's no microbial problems with over-encapsulated products

So, once all the testing is performed, certificate of analysis will be issued--that's a C of A in the slides--so that the product can be now released for the next step. Either it can go directly to the clinic if it's a bulk product that's released or prescribed to the patients, or it can be packaged into various configurations

So, for example, as the slide shows, if it's packaged into blisters or bottles--the reason it's good to have all the bulk products tested before it's blistered or bottled is because, most of the time in the clinical trials, blistering and bottling is done many different configurations. So, you will have blisters with one or two products in it, maybe seven or eight depending on one week supply, one month supply, it may be a one day supply. Same thing with the bottles - many different variety of counts are placed into the bottles

So, if you bottle it blister first and then try to do complete release testing, we might have to test same product ten times. So, it's better to test in bulk. And then, once it's blistered or bottled, just to do the stability testing and the identification testing

Obviously, stability is important for the shelf life determination, but simple identification testing will tell us, yes, we have the right product into the right blisters or right columns of the blisters

And then, individual blisters and bottles can go to the next step. That's the final part of this slide. Blisters can put it into columns or rows in a kit, in a wallet, or different bottles can go into another bigger box to make it a one week supply or a one month supply

Again, identification testing can be performed to ensure the right products are placed into the right location as is intentional--originally intended

As shown here, the stability testing is normally performed on the initial blisters and bottles because that is the point you want to ensure that products are stable. Once it's done into secondary [unintelligible] kits, you don't have to have a stability testing because product's not impacted in any other way

So, as you can imagine, you can have as many as three different types of certificate of analysis, depending on how your products are designed and how your products are packaged. Obviously, if you have only one blister packaging type, we can do all of the release testing such as ID, assay dissolution, microbiology, all on the single blister package

So, I talked about different analytical testing. So, there are basically three big steps that we have to think about. The first is obviously method

Each [unintelligible] products, commercial products have a unique component, unique drugs inside, active components. And therefore, they have to have a unique analytical method that's designed for their specific product. So, we need to have a method

Second one is reference standards. Without the reference standards, we will not know, we will not have anything to compare against. And then, finally, stability studies are required for determining shelf life

So, first, let's talk about the analytical methods. There are many variety of sources that we can find analytical methods for the commercial products. The most common is pharmacopeia. Most commonly--this slides shows US pharmacopeia, normally called USP, or British pharmacopeia has a lot of commercial products that compendium methods already listed there

Oftentimes, though, many of the commercial products that customer wishes to use it as a competitor testing, the products do not have any compendium method because it's so new and there are no genetic products available, so obviously, no compendium methods available. Often, we can find published articles that describes how those products are tested

In addition, every commercial product in the United States already have dissolution methods available in the FDA dissolution database. It describes the medium dissolution method, the time points and when to test it. So, all these are listed in detail

If none of the methods are available, it is possible to develop the methods from scratch to a full method development. So, all of these choices are available

Now, where do you find your reference standard? Again, pharmacopeia, so this USP is the biggest source of reference standard samples. If USP does not have it available, there are many commercial sources available. You can purchase them

Many API suppliers will--happy to sell you five or ten grams of reference standards, or Sigma Aldrich will supply them, as well

In rare cases, we may not be able to find reference standards from any sources because the product is so new to the market. In that case, it is possible to use the commercial product as a reference product and then use that to test all of the blinded product

Obviously, it's a lot more difficult to do so, but it is possible to perform that way

Now, this slide shows a very simple pictorial showing the flow of how analytical test is done such as assays or impurities for those of you who may not be familiar with analytical testing. So, we start with packaged product, remove the products from the packaging, and they are ground with normal [unintelligible]

Once they are completely pulverized [sp] into a white powder, we dissolve it completely and then analyze it, normally instrument called HPLC, and that stands for High Performance Liquid Chromatography. So, this instrument will tell us identity of the drug, how much of drug is in it--so, in other words, potency--as well as all of the impurities that might be present in the product

Now, analytical methods--let's say we have taken the methods from a compendium method such as USP. The compendium method's already validated. However, we still have to do some additional validation

The reason is we are not dealing with original commercial products. We are dealing with blinded product, which is over-encapsulated with a capsule present and with additional backfill in it. So, that might interfere with the analysis itself. So, we have to do some specificity validation

And sometimes, the color of the shell can impact the analytical method, so that has to be also taken into consideration. So, minimum specificity has to be tested and validated for individual existing methods

Now, let me talk about the dissolution testing. I talked about performance of the blinded products, the over-encapsulated products. So, this is how we do the dissolution testing

So, the over-encapsulated products are removed from the packaging, and that's placed into an equipment called dissolution bath. And that's shown on the slide on the right

And normally, we put it into six different identical vessels. It's filled with about a liter of a medium. Normally, it's water or acidic media, depending on how the drug is developed, dissolved in the body

And then, when we stir them or whatever method that's required or specified in the compendium method, the samples are pulled at specific time points - say 15 minutes, 30 minutes, 45 minutes or two hours, depending on the product, and that those pulled samples are again analyzed by HPLC to determine how much is drug is dissolved in the dissolution media within a specified time

There are two normal dissolution apparatus [sp] used for blinded product testing. The one on the left is apparatus one. The one on the right is apparatus two

So, if you look at apparatus two, that's just showing a paddle in the middle. That's the metal piece. And then, it's just sitting inside a round dissolution bath

The dissolution bath is filled with up to the rim almost a liter of the liquid, normally water. And then, as you can see, there's a pink tablet on the bottom, and as the paddle stirs, the tablet dissolves into the dissolution media, and then it releases the drug. And that's how we test, make sure the tablet is dissolving properly, depending on the different product, obviously, different speed and different media is determined

The one on the left is normally used for capsules because tablet, when you put it into the liquid, it sinks to the bottom. Capsules will float

So, if something has to hold a capsule--so, instead of using the paddles, we'll put it into a basket. So, the picture, if you can imagine, this basket is on the place of the paddle on the right inside dissolution bath. So, the capsule is kept inside a basket, and the basket itself is rotating to help to dissolve the drug product inside

Now, if you imagine your blinded product, most of the blinded products start with a tablet. So, the USP method, compendium method will be apparatus two, which requires the paddle

But, because it is a capsule now, you put it into another capsule with a backfill, it will float to the top. So, to be able to use the same apparatus that's in the compendium method, we will use something called sinkers [sp]

So, this slide shows on the bottom right hand slide various types of spiral sinkers and thongs [sp] that holds down the tablet or the capsules to the bottom of the vessel, so when the paddle's spinning, you have your product on the bottom. So, eventually, the capsule will disintegrate and the backfill will dissipate, and then your tablet will be exposed to dissolve inside the vessel

Now, if you already have capsules starting as a commercial product, then you make a blinded product, obviously, you'll be using the same basket. So, in the picture in the slide, there are various types of baskets. They can be used for different various products

So, to continue this talk about the dissolution techniques, the reason dissolution is such an important technique for determining the performance of your blinded product is shown on this slide. Because you have taken the commercial product and put it inside another capsule shell with a backfill, that oftentimes delay the dissolution

So, if your over-encapsulated product dissolves slower than the original commercial product, you may have a failure and you may have questions with the bio equivalence. So, because of that, oftentimes, we will do comparative dissolution, which I'll talk about in the next slides

So, in this slide, there is a very nice diagram of dissolution profile. So, if you look at the X axis, that's the time points. The Y axis shows percent dissolved. So, the dissolution results are shown from zero all the way to the 100 percent

And the dark blue line on the top, that profile is the original commercial tablet. So, it's dissolving, as you can see, about 60 minutes, about 70 percent dissolved, at 120 minutes, it's over 80 percent dissolved

The pink line on the bottom is over-encapsulated tablets. So, that's your blinded material. As you can see already immediately, the pink line is much lower in terms of dissolution than the commercial product so that you already have delayed dissolution

So, the question is, is this a bioequivalent. And when we test that, we answer that question is this a bioequivalent by calculating something called similarity factor. That's shown on this slide

So, in order to calculate similarity factor, we perform the dissolution with [unintelligible] 12. That means you perform the dissolution of 12 original tablets, original product, and 12 of the blinded over-encapsulated product

And after the dissolution testing is completed, something called similarity factor is calculated, the equations are shown here. And if the answer is between 50 to 100, that indicates two products are similar, and that passes the similarity factor, that means the over-encapsulated products are performing similarly to the commercial products. So, it can be released to the clinic

Now, here is an example, again showing two dissolution profiles with original tablet, and then the pink line on the bottom is the over-encapsulated blinded product

As we can see, the pink line is not dissolving very well at all. So, if you look at time points such as 40, 40 minutes, about 40 and 50 minutes, the original product dissolves almost 100 percent whereas the over-encapsulated product, the pink line is dissolved only about 60 percent

The reason is obvious. If you look at the picture on the right shows bottom of dissolution vessel

So, what this is shown is that the capsule that's outside has already disintegrated. But, inside the capsule, you have a backfill, which is normally excipient

Now, most excipient used for backfill are polymers. So, they're not water soluble

So, if you have very small tablet and use large quantities of backfill, the backfill will sit right on top of your tablet, and that's what we call coning [sp]. When it's coning, no matter how much the paddle is spinning above it, the excipient might be just happily sit there on top of your tablet. And when that happens, the drug is not released because it's covered inside a mound of your backfill, which is the polymer excipient

So, as you can see from the graph from the dissolution profiles that the over-encapsulated product is very slowly dissolving. Even after almost between 40 and 50 minutes, it's really released about 60 percent. And only after 60 minutes, it's released more than 80 percent

So, let's assume if this product specification was 80 percent dissolved after 60 minutes, this product will pass. The over-encapsulated product will also pass the specification. But, this will fail the similarity factor

That's why it's important to know, not only meeting the specification of dissolution percent dissolved, but the two different profiles of the unmodified product and the blinded product has to have a similar dissolution profile to pass it. That's why similarity factor, which is done by comparative dissolution, and normal specification of how much to dissolve have to both pass

Now--so, what do you do if this happens to your own product? Well, there are two ways to solve this problem. Obviously, the easiest way is change your backfill

If you change to different excipient, oftentimes, finer smaller particles will dissipate far easier, and that will solve the problem. However, oftentimes, it might be too late to change excipient because the product may have been already manufactured. In that case, we'll have to modify the dissolution method

So, once the dissolution method is modified, it has to be validated. And then, comparative dissolution has to be repeated against a modified, the commercial product, and the blinded material using the identical modified method to show that the similarity factor is passing and everything is dissolving in a similar way

The next slide shows a different kind of problem. It's something called cross-linking

Because over-encapsulation is normally happening with a hard gelatin capsule, the gelatin material can, over time, cross link and becomes very hard to disintegrate. Normal gelatin capsule will disintegrate in water in about five minutes

When they cross-link, they may not want to disintegrate at all and might hold onto it for a very long time, which will cause failure on the dissolution testing

The reason it's happening is because, sometimes, your product, your product, commercial product itself, or the backfill might react with gelatin capsule because they are not compatible, and that might cause cross-linking. Oftentimes, the true product tablets that are coated with the various materials, the coating can cause cross-linking of the gelatin capsules. And sometimes, the solubility is decreased because of the storage at high temperature and high humidity

So, if you look at the right hand side picture, again, this is showing a bottom of the dissolution vessel, and you can see there's a pink capsule--no, it's blue capsules inside, not pink, blue capsules inside. And as you can see, the capsule is held up by a spiral stainless steel sinker. So, you can see the sinker on the picture

Now, normally, when you put the capsules in the media, within five minutes, capsules will disintegrate, and then the--you will immediately see the backfill and the tablets inside

In this case, even after long time of storing, instead of capsules disintegrating, it absorbs media and becomes bigger and bigger and turns into a big sponge. And then, it doesn't release the product

Now, that's evident from the left hand side where the dissolution profile is shown. Again, X axis is tie [sp], Y axis is percent dissolved. The dark blue line on the top is a tablet. And it's dissolved about, after 30 minutes, almost 100 percent dissolved

And then, the red line under it is the blinded material, which is over-encapsulated tablet. And that gives a very good profile, as well - at 30 minutes, close to 100 percent is dissolved, about 90 percent dissolved. So, it looks really good

But, if you look at the bottom graph, which is green, that's your blinded product, over-encapsulated product. After one month, that's one--M stands as one month--40/75--we'll talk about the stability study later, but 40/75 indicates it was stored for one month at 40 degrees C, 75 percent humidity

So, after one month, as you can see the dissolution profile, for 30, first 30 minutes, no drugs dissolved. Obviously, the capsule shell is expanding, expanding. It's not releasing any of the drug

And then, about 45 minutes, you see a little bit of drug is dissolved, about 30 percent dissolved. And 60 minutes, another 40 percent is dissolved

Obviously, if this happens, this product will fail after one month on stability

So, when this happens, there are other ways to overcome it. USP allows two different types of enzymes to be used to basically digest the capsule shells when it's cross-linked and not dissolving, and those are pepsin and pancreatic

So, depending on the PH of the dissolution media, you can choose one of the enzymes to help digest the capsule shells because those enzymes are obviously present in our body. And so, you can successfully complete the dissolution testing during stability studies

So, we've been talking about the stability and some additional things that we have to think about, because every over-encapsulated blinded product has to be placed on the stability study to determine the new shelf life. Again, we have to think about excipient interactions and the capsule shell itself because these two can have a big impact on the stability of a blinded material

Excipients are normally selected by what's already in the tablet. So, if the tablet contains specific excipient, normally, that same excipient is selected for over-encapsulation. However, that doesn't always guarantee the stability. Sometimes, additional excipient can impact the stability of the product

Now, capsule shells - the biggest drawback of capsule shells is that it has 11 percent moisture. So, if the true product is very moisture sensitive, they can interact with the capsule shell and interfere with the stability and even increase the degradation rate. And sometimes, capsule shell itself, the material itself may not be comparable with the API itself

So, there are different types of capsule shells. Instead of gelatin capsule shells, there are vegetable capsule shells with a different moisture content

Or sometimes, a customer will overcome the moisture problems by adding additional drying cylinders inside containers such as [unintelligible] into the bottles to avoid having API degradation

So, to perform the stability study, we normally advise to place not only your modified, the over-encapsulated blinded products into the stability, but also put the original commercial product in its original packaging in the same condition and the placebo products. The reason you want to do that is, sometimes, your blinded product may fail stability studies, but it may not be because of blinding itself. It might be because the original product is unstable

So, you want to always eliminate the question why it's failing. Maybe it's original question. So, that has to be always done concurrently

You don't have to test both products at every time point, but have it available so that if your blinded material fails the stability study, then you can go back to the original commercial product that's placed in the same condition and analyze that to see where the failure came from

Obviously, you want to test placebo even though we are only doing visual testing for placebo because you don't want to have placebo inactive [sp] changing to different colors for any blinded studies

Now, if you have a multiple strength, because a lot of clinical trials will have more than three to five strengths tested for blinding test, then bracketing can be used as long as the formulation is consistent between the low strengths to high strength

Some of the delayed release or the extended release tablets, we may not be able to do bracketing studies because of different strength may have a quite different formulations. So, all of them may have to be tested for stability

And I've listed four different conditions, and these are very common conditions for stability studies. So, for example, for North America and most of Europe, the normal storage conditions are climactic zone two, that's 25 degrees C, 60 percent RH

So, the product will be placed in that stability condition to monitor during the duration of our clinical trials. At the same time, products are placed in something called accelerated conditions. That’s on the bottom list. That's 40 degrees C, 75 percent RH

So, depending on your climactic zone, you have your zone temperature and then the accelerated conditions

So, there is more details of how the stability study is done on this current slide, and it shows all the different time points. So, this is example of North America and most of Europe where the climactic zone is two. So, our standard condition is 25 degrees C, 60 percent RH

The testing frequency is--required is every three months up to one year--so, it's three, six, nine, 12 months--and then every six months after two years--so, 12, 18, 24. And after that, it's annual testing if it's required to be stored longer

And then, on the bottom, there's accelerated condition. That's 40 degrees C, 75 percent humidity. And that's tested only up to six months

Now, normally, it's three months and six months testing, but most customers prefer testing at one month because one month data gives you a lot of information how stable this product would be

We also recommend putting samples in 30 degrees C, 65 percent RH. The reason is because some products might be unstable at 40 but might be stable at 30. So, if it fails at 40 degrees C, accelerated conditions, we test up to one year at 30 degrees C, 65 percent humidity to confirm--to have that as accelerated conditions

Obviously, if the commercial product is stored at refrigerated conditions, then refrigerated condition would be the long term and 25 degrees C, 60 percent will be the accelerated. So, as you can see, depending on where the current drug storage condition is, we have to determine what the accelerated condition is

So, we talked about the stability study. So, what do we do at each time point? The stability testing time point, the very important first thing, visual inspection. We make sure the capsules didn't change in color, shape, the backfill didn't discolor it and the tablets inside didn't change visually

And then, what's the point of doing stability testing? It's because you want to make sure the product is stable over the period of time

Therefore, we always do assay. That's to determine the potency, so the drug's not decreasing in potency, and the related substances. That's to determine that impurity's not increasing

If the impurity's increasing above and beyond over what's specified for the product to be acceptable, if there is increase, that means the stability study's over. You can no longer use that sample. And that will determine the shelf life of that sample as up to that point

Obviously, dissolution testing has to be always included because dissolution testing determines--ensures that the blinded over-encapsulated products are not changing in terms of performance during the storage, during the duration of your shelf life and the clinical trials

Now, moisture is tested on some of the products, especially if you know this commercial product is moisture sensitive. Then, moisture may be tested at each time point, make sure that there is no change of the true product and no moisture is taken in from the environment

So, if I summarize all the process that we discussed today, the very first step for performing all of the analytical testing for the blinded products is analytical methods, either from literature, compendium method, whichever source have to have analytical method. And then, secondly, we need to have a reference standard material. So, I talked about where to get those reference standard materials

And also, there is some developing analytical methods, some validation is always involved because it's over-encapsulated product. Therefore, it's not the same as a commercial product, so compendium method cannot be used verbatim. It has to be always validated, make sure it works for these modified blinded product, as well

Now, comparative dissolution - I talked about how important it is to determine the performance did not change and to confirm the blinded products, the over-encapsulated products are bio equivalent to the commercial product, we have to perform the comparative dissolution

Finally, stability study we discussed because during the duration of your clinical trials, you want to make sure that you have sufficient shelf life to use the same lot, same lot of product continuously

Now, I will talk about, something about the timelines. As you can imagine, all these analytical methods, validations, some upfront work, we like to have at least three months of stability data before starting the clinical trials because you don't want to have any surprises on your huge clinical batch and have an unexpected failure

So, it is a very common practice to make a very small batch of stability batch just to solely purpose of testing for stability data only. So, it's a fairly small batch, even though it's manufactured identical way as a clinical batch would. So, you do that ahead of time and obtain three month stability data, and then that gives the confidence that the clinical batch would go fairly well and you'll have no surprises or problems during the clinical trials

So, to discuss clinical trial materials, this means you have to start the discussion at least six months before that because you want to have at least three months stability data

Now, I will briefly talk about different types of testing required at each different time point. So, first, let's talk about release testing, what type of testing do we have to do

For over-encapsulated material, obviously, you need to have a specification indicating what is a pass fail limit. Comparative dissolution data is important for the release testing. That's most important. That's the minimum important you need to have, absolute minimum

In addition to that, oftentimes, assay and related substance are included into the release testing. The reason is because you want to compare the data to the stability data later

Appearance, obviously, is always included, and at the end, a certificate of analysis released to release a product into the clinic

I have an example of a placebo inhaler devices because, you know, inhaler devices are often tested, but placebo is not often tested. So, I'd like to talk about it a little bit because it is important to test placebos

Obviously, same story - you need to have analytical method. And specification has to be set

Now, the placebo devices can be tested in two different ways. We can test disassembled device, or we can actually test the immediate dose. The reason is we want to make sure there is no active present in the placebo device, and the certificate of analysis is issued

Now, wallets [sp] will have been already tested before it's put into wallets, either it's in a blister stage or bulk stage. But, once put it into wallets, it's oftentimes very important to do additional ID testing, that's identification testing. The reason is wallet designs are very oftentimes complex, and you want to make sure the right dose and right product is in the right column or right row, because sometimes, you'll have different strength or active and placebo mixed in the same wallet cards

So, obviously, method, specification has to be all present before that

Assay method - that's to determine strength if you have different doses inside same wallet cards. And certificate of analysis will be issued

So, I have next slide an example of a wallet. This is a fairly extreme case that we have. But, as you can see, there is a morning column and the evening column. And there are various different strength put into the wallet cards

So, if you have different strength, unless individual capsules can be opened and we can identify it by looking at visually what strength of tablets they're in--sometimes, we can focal measure products--we have to do actual assay testing to determine the right product is in the right location of the wallet card

I have two case studies, just to give you a little bit of flavor for how this clinical trial product manufacturing goes from the beginning to the point of clinical batch production begins

So, in this first case study one, we studied discussion and planning--obviously, month one from the beginning. And then, month two, we did a feasibility study. What does a feasibility study mean? We don't know if this commercial product can be put into certain capsules, and then what type of excipients do you use, and will it meet the dissolution parameters, will it pass the performance testing

So, to do all of that, we did initial feasibility study. To do that, it's very simple. You make man encapsulations in the lab. You can make 50 to 100 blinded products by hand. And then, we will test them to make sure performance is not impacted. You can even test various different excipients, make sure you can select the right excipient. You can use the same capsules for validation studies

Once that study's done, obviously, we already know what capsules to use, what excipient to use. So, month three, stability batch can be produced. That stability batch is that the over-capsulated product that's put into either bottles or some type of blister packages

While the stability batch is under production, then analytical lab will perform validation studies to make sure this over-encapsulated--over-encapsulation process is not impacting actual analytical test, and then the methods are also specific for the product

Month four, the stability batch production was completed. So, we were able to put all of the products into the stability conditions on the various conditions

Now, in this case, we only did up to six months stability studies. It's depending on the customer. Some customers wish to do full 24 month stability study, even on the stability batch, not on the clinical batch, but the stability batch, because it always gives them three months data before they get the data for the clinical batch. So, it gives a nice advanced notice if something might go wrong

However, some customers have to do a short term stability study on stability batch, and in this case, we did actually three months stability studies with a six months as optional. If we see something funny or something concerned at three months, we might do another six months poll

The next follow month, month five, we did a one month stability testing. Everything looked okay. And then, so since everything looked good, clinical batch production was scheduled

And then, month seven, we were able to do three months stability testing. Now, remember this is both conditions, 25 degrees C, 60 percent RH, which is normal conditions, and the accelerated conditions, that's 40 degrees C, 75 percent RH

So, at both conditions, we test at three months, and that gave confidence to the customer to give a green light to start the clinical batch production

So, the whole process took about seven months to begin the production

Now, next, this is the final slide for case study, second case study. This one was a little bit shorter because we took a little bit of risk

So, you can see from these timelines that it is possible to even make the timeline even shorter, depending on how much risk you want to--you're willing to take

So, at first month, we started discussion and the planning of the analytical testing. At month two, we studied dissolution testing and then start validating all the methods

But, at the same time, stability batch production began. So, stability batch production started before we confirmed that this product will have possible or acceptable performance once we blinded, because that was assumed that it'll work

And so, month three, as soon as stability batch was produced, we started stability studies. Now, this case, we had 24 month stability study

Month four--so, by that time, month four arrived, we had all of the methods validated. So, we did T Zero [sp] testing, which is initial testing, and one month testing all together at month four using the stability batch

So, obviously, one month testing is from the samples that already in stability condition, the accelerated condition. T Zero testing is the samples, the original samples that we have not put it into the stability conditions yet

Obviously, we can do this for various reasons. First of all, we knew that the customer's taking the risk because the timeline was shorter. And secondly, this is a stability batch, not the clinical batch. Therefore, we can test times zero point and one month point at the same time

Now, one month data was good. There was nothing concerning. And then, six months time point, we were able to perform three month stability testing so that clinical batch production was able to start at the end of six months

Now, it is possible for some customers to skip stability batch production entirely. There are multiple ways--multiple reasons why you can do that

Obviously, one, sometimes, the timeline is so tight, because you only have two months to discuss and you don't have time to make stability batches and clinical trial batches. However, because companies like us, if you come to service companies like us, because we have performed hundreds of stability studies in the past, we might have already performed stability study on the same commercial product that you are wishing to test

It may not be the same manufacturers, but it might be the same product. And therefore, we might have already advanced knowledge or information to tell you that, yes, we have test this product with this configuration and it was stable up to two years. And that might give you confidence, even though you have very little time, to start immediately, to start clinical batch production without going through all the stability batch production

In some cases, some products are known to be very, very stable. And those cases, oftentimes, customers will skip stability batch production and go straight to the clinical batch production

So, today, I talked about all the important points of testing, blinded, over-encapsulated clinical products. So, those testing requirements, you have to have a method. And I talked about how to get those method

And I also talked about what type of parameters have to be validated at minimum. And I talked about how to get the reference standard materials and talked about importance of performance testing using dissolution and comparative dissolutions and also talked about some of the issues that we have encountered during the dissolution and how you can overcome that and make improvements

And finally, I talked about the stability testing, the importance of it and what type of things that has to be tested for stability studies

And thank you for your time, and if you have any questions, please feel free to contact any of us Almac. Thank you. Have a good day