Ultralente crystal size

too much time on your hands, Jim?

dave

Jim Dumas said:

I've been reading Novo patents for crystal sizes. The most
promising US patent is #2799622: "Process of Producing
Insulin Crystals of Substantially Uniform Size and
Compositions Thereof," 16 July, 1957.

This patent provides an equation to estimate seed crystal
requirements given seed crystal size, required finished
crystal size and weight of insulin that will be
crystalized. The examples focus on 30 micron required
finished size and states it is possible to obtain 80-90%
in the form of 28-36 micron crystals. This diameter is the
longest diagonal distance across the crystals. Assuming a
normal (Gaussian) distribution, setting the mean to 32
microns and forcing the difference of integrals:

int(28,infinity) - int(36, infinity) = 0.85 or 85% yield

This gives a standard deviation of 2.75 microns in crystal
size. So the mean+2SD=~37.5 or about 38 microns for the
maximum crystal size in commercial ultralente. The lower
size is probably 26 microns.

This assumes a perfect seed size of 4 microns as
recommended in the patent. If the manufacturer is sloppy
then the sizes could be less uniform.

In any case, this is very close to Michel's 40
micron number.

This range of crystal sizes in ultralente could cause
variable subcutaneous absorption characteristics. It
also suggests there could be differences in crystal
sizes between batches from the manufacturer (only Lilly
these days).

Novo patent US#2882202: "Insulin Crystal Preparations and
Methods of Producting Them," 14 April, 1959; states 10
micron crystals produce the same basal insulinemia effect
(time course of action) as amorphous PZI in dog models.
Crystal sizes of 25-100 microns have longer effects than
PZI in dog models. This is an excellent benchmark to keep
in mind as it probably set the goal for the 30 micron
design size above.

Food for thought,

Bay Area Dave said:

too much time on your hands, Jim?

Not as much as you obviously have.
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

how do you figure? my posts are short and to the point.
little time spent! 🙂

Jim Dumas said:

Bay Area Dave said:

too much time on your hands, Jim?

Not as much as you obviously have.

Bay Area Dave said:

how do you figure?

Easy. Your post was designed to cause trouble. Therefore,
you have much more time on your hands than I do.

Two years ago I posted to MHD that I was thinking about
making my own ultralente rDNA insulin if Lilly discontinued
UL (Google on "Michel: How to make UL?"😉. These patents
describe various methods to produce insulin crystals and
are in keeping with my interest in this area. Michel Martin
had successfully done this with beef insulin but the
equipment required was expensive. I think I can do this on
the cheap. So the patent search is necessary to understand
the prior art.

In any case, I figure you're not interested UL and
should nose out.
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

cause trouble?? my, my you are awful touchy, Jim. neither of
my 2 previous posts were rude. I was just teasing you.
apparently you aren't amenable to teasing. I shall leave you
to your studies and shall try to refrain from engaging you
in the future.

in fact, let's make it for sure--PLONK!

Feel better?

dave

Jim Dumas said:

Bay Area Dave said:

how do you figure?

Easy. Your post was designed to cause trouble. Therefore,
you have much more time on your hands than I do.

Two years ago I posted to MHD that I was thinking about
making my own ultralente rDNA insulin if Lilly
discontinued UL (Google on "Michel: How to make UL?"😉.
These patents describe various methods to produce insulin
crystals and are in keeping with my interest in this area.
Michel Martin had successfully done this with beef insulin
but the equipment required was expensive. I think I can do
this on the cheap. So the patent search is necessary to
understand the prior art.

In any case, I figure you're not interested UL and should
nose out.

Bay Area Dave said:

Feel better?

Indeed I do. Thank God for minor miracles.
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

Hey Dave, do not mess with the animal insulin people.

Most of them are technically competent. Some people find
that animal insulin works better for them. There is a
serious threat to the supply.

I found It did not work better for me but all diabetics are
not the same. Jim Dumas has been an excellent source of
information for me. I can say the same for Michael Martin.
There are more issue to diabetes that the common items. Live
and let live.

I need all of the help I can get. Including you. Guy

On Thu, 08 Apr 2004 01:53:38 GMT, Jim Dumas <j-

[email protected]!mindspring.com said:

Bay Area Dave said:

Feel better?

Indeed I do. Thank God for minor miracles.

I wouldn't (and didn't) say that Jim didn't know his
"technical stuff". I was commenting on the fact that he
posted something that is over the heads of us mere
mortals, but I didn't do it in a disrespectfully way. He
took offense, unfortunately. Since he argued
unsuccessfully with me ad infinitum in the recent past, I
find it relevant to shield his posts from my eyes in order
not to clutter the NG with endless arguing over fine
points that neither will concede.

My intention is to limit endless debate that won't solve
anything. my opinion vs his opinion. it's fruitless.

doesn't that sound reasonable to you, Guy?

dave

Guy said:

Hey Dave, do not mess with the animal insulin people.

Most of them are technically competent. Some people find
that animal insulin works better for them. There is a
serious threat to the supply.

I found It did not work better for me but all diabetics
are not the same. Jim Dumas has been an excellent source
of information for me. I can say the same for Michael
Martin. There are more issue to diabetes that the common
items. Live and let live.

I need all of the help I can get. Including you. Guy

On Thu, 08 Apr 2004 01:53:38 GMT, Jim Dumas <j-

[email protected]!mindspring.com said:

Bay Area Dave said:

Feel better?

Indeed I do. Thank God for minor miracles.

Bay Area Dave <[email hidden]> wrote on Thu, 08 Apr 2004 03:32:17 GMT:

Quoted message said:

I wouldn't (and didn't) say that Jim didn't know his
"technical stuff". I was commenting on the fact that he
posted something that is over the heads of us mere
mortals, but I didn't do it in a disrespectfully way. He
took offense, unfortunately.

I found your first post disrespectful. Playfully
intended perhaps, but somewhat ambiguous in tone. Jim's
reply, objecting to that post was fully in order - not
the only proper way to take your post, but a valid one,
none the less.

Your _second_ post, confirmed and anchored the disrespect
which was only latent in your first, whatever the intent of
that first might have been. A short, even lighthearted
apology wouldn't have been out of place there.

Quoted message said:

Since he argued unsuccessfully with me ad infinitum in the
recent past, I find it relevant to shield his posts from
my eyes in order not to clutter the NG with endless
arguing over fine points that neither will concede.

Quoted message said:

My intention is to limit endless debate that won't solve
anything. my opinion vs his opinion. it's fruitless.

However, your action seems almost optimally designed for the
prolongation of bitter squabling.

Let's not do this here. Please.

Quoted message said:

dave

--
Alan Mackenzie (Munich, Germany) Email: [email hidden]; to
decode, wherever there is a repeated letter (like "aa"😉,
remove half of them (leaving, say, "a"😉.

In article <[email hidden]>,

Bay Area Dave said:

I wouldn't (and didn't) say that Jim didn't know his
"technical stuff". I was commenting on the fact that he
posted something that is over the heads of us mere mortals,

It should not be over the head of anyone with a good enough
education to be able to make intelligent decisions.

--
This address is for information only. I do not claim that
these views are those of the Statistics Department or of
Purdue University. Herman Rubin, Department of Statistics,
Purdue University [email hidden] Phone: (765)494-
6054 FAX: (765)494-0558

this thread would have ended last night had you not
interjected. while I appreciate your psychoanalytical
skills, this thread is finito. Jim's response was uncalled
for with respect to my FIRST post. we can either agree to
disagree OR you can argue with yourself, as I'm D O N E
arguing this thread.

thanks for your cooperation.

dave

Alan Mackenzie said:

Bay Area Dave <[email hidden]> wrote on Thu, 08 Apr 2004
03:32:17 GMT:

Quoted message said:

I wouldn't (and didn't) say that Jim didn't know his
"technical stuff". I was commenting on the fact that he
posted something that is over the heads of us mere
mortals, but I didn't do it in a disrespectfully way. He
took offense, unfortunately.

I found your first post disrespectful. Playfully
intended perhaps, but somewhat ambiguous in tone. Jim's
reply, objecting to that post was fully in order - not
the only proper way to take your post, but a valid one,
none the less.

Your _second_ post, confirmed and anchored the disrespect
which was only latent in your first, whatever the intent
of that first might have been. A short, even lighthearted
apology wouldn't have been out of place there.

Quoted message said:

Since he argued unsuccessfully with me ad infinitum in the
recent past, I find it relevant to shield his posts from
my eyes in order not to clutter the NG with endless
arguing over fine points that neither will concede.

Quoted message said:

My intention is to limit endless debate that won't solve
anything. my opinion vs his opinion. it's fruitless.

However, your action seems almost optimally designed for
the prolongation of bitter squabling.

Let's not do this here. Please.

Quoted message said:

dave

Jim Dumas said:

I've been reading Novo patents for crystal sizes. The most
promising US patent is #2799622: "Process of Producing
Insulin Crystals of Substantially Uniform Size and
Compositions Thereof," 16 July, 1957.

This patent is an ultralente gold mine. The time to
crystallize with constant stirring is 15-20 hours for seed
crystals of 1-7 microns. It also states the design goal is
for crystal sizes from 10-40 microns.

So I'm revising my design requirements to a mean of 30
microns. This gives a 2 micron standard deviation for the
integral difference of:

Gaussian integral(28 to infinity) - integral(36 to
infinity) = 0.84

or an 84% yield (80-90% req'd) for crystal sizes of 28-
36 microns.

Next, I will not filter my preparation but will accept
Gaussian tails on both sides of the 30 micron average. This
should produce 95% of the crystals between 26-34 microns
(mean +/- 2SD). The integral difference for crystals between
10 and 40 microns is 0.9999997! That is, most crystals are
the correct size to give prolonged subcutaneous absorption
without filtering small or large crystals out. The integral
for 40 to infinity is
2.87e-7 so there should be no crystals larger than 40
microns in the preparation.

The tricky part is getting the optimum 4 micron seed
crystals. The next issue is the pH (5-7 required for
crystallization). It will probably drop to 6 with the
high zinc concentration. But Lantus is an acid pH so I
don't think this is an issue for injecting subcutaneously
(pain-wise).

The basic idea is to take a 10 ml vial of rDNA Humulin R,
add the correct number of sterile 4 micron seed crystals and
zinc salt. Then use my BBQ rotor to tumble the mixture for a
day at room temperature. Then remove excess fluid to achieve
U-100 again. The zinc content may also have to be lowered by
the same removal then add sterile distilled/deionized water.
This will also bring the pH up closer to neutral. But I
think this is possible with little expense in equipment.

So how to get/make 4 micron seed crystals is the show
stopper.
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

On Thu, 08 Apr 2004 19:10:54 GMT, Jim Dumas

[email protected]!mindspring.com said:

The basic idea is to take a 10 ml vial of rDNA Humulin R,
add the correct number of sterile 4 micron seed crystals
and zinc salt. Then use my BBQ rotor to tumble the mixture
for a day at room temperature. Then remove excess fluid to
achieve U-100 again. The zinc content may also have to be
lowered by the same removal then add sterile
distilled/deionized water. This will also bring the pH up
closer to neutral. But I think this is possible with little
expense in equipment.

So how to get/make 4 micron seed crystals is the show
stopper.

Are you actually considering making your own insulin? It's
fun to play with statistics and bell curves, but making
insulin is next to crazy. Suppose you make it, who will be
the guinea pig for the first injection?

_____________________________________________
tcainternet.comindex.htmlOpen ↗ Free
Diabetic Software

Jim Dumas said:

I've been reading Novo patents for crystal sizes. The most
promising US patent is #2799622: "Process of Producing
Insulin Crystals of Substantially Uniform Size and
Compositions Thereof," 16 July, 1957.

This patent provides an equation to estimate seed crystal
requirements given seed crystal size, required finished
crystal size and weight of insulin that will be
crystalized. The examples focus on 30 micron required
finished size and states it is possible to obtain 80-90%
in the form of 28-36 micron crystals. This diameter is the
longest diagonal distance across the crystals. Assuming a
normal (Gaussian) distribution, setting the mean to 32
microns and forcing the difference of integrals:

<snip>

Quoted message said:

In any case, this is very close to Michel's 40
micron number.

This range of crystal sizes in ultralente could cause
variable subcutaneous absorption characteristics. It
also suggests there could be differences in crystal
sizes between batches from the manufacturer (only Lilly
these days).

<snip>

another thought...

almost everyone seems to think of Lente as an
"intermediate" insulin

nothing could be further from the truth!!!

Lente is 70% UL

there are at least 3 different companies (on the world
market) who are still making various Lente insulins. 🙂

if Novo still does Lente (i.e. "human"-L), that'd make 4

bill t1 since '57, ex 8-yr pumper, pork/beef-L 2x,
simple MDI/DAFNE

Jim Dumas said:

Two years ago I posted to MHD that I was thinking about
making my own ultralente rDNA insulin if Lilly
discontinued UL (Google on "Michel: How to make UL?"😉.
These patents describe various methods to produce insulin
crystals and are in keeping with my interest in this area.
Michel Martin had successfully done this with beef insulin

incorrect

mickey found a way to make pork-SL (SemiLente) from pork-R

he called it "quasi-Lente" 🙂

fwiw, Novo's pork-R gave him better results than
Lilly's pork-R

i'd suggest you e-mail him

this assumes that he's still alive now that he's on rDNA
[censored]

bill t1 since '57, pork/beef-L 2x, simple MDI/DAFNE

Quoted message said:

but the equipment required was expensive. I think I can do
this on the cheap. So the patent search is necessary to
understand the prior art.

In any case, I figure you're not interested UL and should
nose out.

Hi_Therre said:

Are you actually considering making your own insulin? It's
fun to play with statistics and bell curves, but making
insulin is next to crazy. Suppose you make it, who will be
the guinea pig for the first injection?

The issue is the loss of human rDNA ultralente from the US
market. This has been expected for two years now. Novo
discontinued human ultralente in the US a few years ago, but
marketed it in Canada until recently (I think it may be
discontinued in Canada, but not sure). My therapeutic goal
is to stay with the human insulin molecule until hell
freezes over. The only analogue I plan to use is Humalog,
since it has not provoked an immune response from my body,
(glucose disposal peaks in 1-2 hours as advertised by Lilly
so no antibody binding delays).

If UL is eventually discontinued in the US, then I will try
to make my own for personal use. I plan to be the test
subject and would probably buy a cheap microscope to do a QC
check on crystal sizes for each batch.

Note that Lilly mentions the possibility of adding zinc to
current human rDNA ultralente insulin, to achieve longer
action than beef ultralente. This was in patent number
5534488 where some kit would be sold to pharmacies to mix at
the pharmacy for patients that require longer insulin action
than current UL provides. So this idea of mixing on demand
has been explored by Lilly, at least.

In my case, the kit would also include 4 micron seed
crystals along with the zinc. Then slowly tumble it for 24
hours to form the larger crystals. All I'm doing is taking
the R hexamers and forming large crystals. So I'm not making
the rDNA insulin, per se. I'm just recrystallizing it.

And I don't think it's hard to do. But still researching
the method,
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

willbill said:

incorrect

mickey found a way to make pork-SL (SemiLente) from pork-R

he called it "quasi-Lente" 🙂

fwiw, Novo's pork-R gave him better results than
Lilly's pork-R

i'd suggest you e-mail him

Thanks Bill,

So Michel didn't find a way to make seed crystals. He added
zinc without seed crystals and SL resulted.

Good info. Guess I'll email him. Thanks,
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

willbill said:

Lente is 70% UL

there are at least 3 different companies (on the world
market) who are still making various Lente insulins. 🙂

Agreed.

Lente is a good substitute for UL. So this is another
avenue. We have:

CP Pharma Lilly USV-India

Not sure about Novo's L.

But if the R->UL works, then the FDA is out of the loop and
the supplies are local. So this has merit in a pinch. It
assumes R will always be available. This could be a problem
as analogues continue to be developed.

Lente is a good suggestion. Thanks,
--
Jim Dumas T1 4/86, background retinopathy, rarely
hypoglycemic: <1/mo. lispro+R+U+NPH daily, moderate
exercise, typically <6% HbA1c

On Fri, 09 Apr 2004 01:01:23 GMT, Jim Dumas

[email protected]!mindspring.com said:

Lente is a good suggestion. Thanks,

Jim,has any work been done on a capsule of insulin under
pressure being fed through a capillary. I have seen
applications where this principle was used for other things.
With a long capillary the flow rate is almost constant over
a wide range of pressures.

Maybe a basal for a week before inserting of a new capsule.

Or even go wilder and find a membrane that would respond to
blood sugars. I would to hear about the results of any work
in these areas.

Guy