Isodiazepam: naturally occurring, novel benzodiazepine

[wungchow]

As far as I know, it's never been marketed...

Do you think this would retain activity?

 

[wungchow]

There's one reference I could find, but I have no access:

EJMCA5 European Journal of Medicinal Chemistry--Chimie Therapeutique.
(Editions Scientifiques Elsevier, 29 rue Buffon, F-75005, Paris, France)
V.9- 1974- Volume(issue)/page/year: 11,83,1976

 

[Phener]

naturally occuring? Would need STRONG evidence for that please, in particular look for g/g of active. [isodiazepam is a metabolite of diazepam] The ability of HPLC-massspec to pick up things is getting TOO good

Benzodiazepines (and DIAZEPAM) have to be one of the most widely used drug and hence pollutant in waste water: Depending on quantities it is more than plausible that specific plants, humans, everything absorb through recycling water. To just emphasize the point

Detection of desmethyldiazepam and diazepam in brain of different species and plants
by
Unseld E, Krishna DR, Fischer C, Klotz U.
Dr. Margarete Fischer-Bosch-Institut für Klinische Pharmakologie,
Stuttgart, Federal Republic of Germany.
Biochem Pharmacol. 1989 Aug 1;3815):2473-8.

ABSTRACT
Recent data suggest that desmethyldiazepam (DD), a major metabolite of several benzodiazepines (BZD), might be of natural origin. Therefore we tried to quantify DD and diazepam (D) in animals during maturation (e.g. hen, chicken, eggs), in brain of species at different evolutionary stages e.g. salmon, frog, monitor/reptile, rat, cat, dog, deer, bovine) including newborn and adult humans. Since low concentrations of DD (range 0.01-0.04 ng/g wet wt) and D (range 0.005-0.02 ng/g) could be measured in different species by sensitive and specific mass spectrometry (GC-MS), we analysed also several plants (e.g. maize corn, lentils, potatoes, soybeans, rice, mushrooms). Again, DD and D could be detected in low amounts (0.005-0.05 ng/g) in some plant products. This would suggest that DD and D might be of natural origin and incorporated via the foodchain into the animal and human body. The biological role or clinical relevance of these intriguing findings need still to be elucidated.

0.05ng/g = 0.00005micrograms/g = 0.00000005mg/g = 0.0000000005g/g

5mg in 20,000,000kg

naturally occuring OR natural polutant from the BUCKETS of diazepam dispensed, eaten and excreted (+/- metabolites) in waste water

Occurrence of “natural” benzodiazepines




References and further reading may be available for this article. To view references and further reading you must purchase this article.


Ulrich Klotza

aDr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology, Stuttgart/Germany

Received 5 November 1990. Available online 16 November 2002.

Abstract
There is accumulating evidence that benzodiazepines (BZD) — agents widely used as anxiolytics and hypnotics-could be regarded as “natural” drugs since they have been found in trace amounts also in plants, various tissues of different animal species and even humans. The biosynthesis of such BZD is still unknown and the hypothesis is favoured that they may be of plant origin. Besides diazepam (D) and its major metabolite desmethyldiazepam (DD) several other BZD (e.g. delorazepam, deschloro-diazepam, delormetazepam, isodiazepam, lormetazepam, oxazepam) could be detected. In some cases identification of these compounds was accomplished by specific mass spectrometry (GC-MS) and for quantification various methods have been applied resulting in different concentrations which range for D from about 0.005 to 1 ng/g and for DD from 0.01 to 0.5 ng/g. It is very unlikely that these trace amounts exert any direct pharmacological effects and at the moment only speculations upon their physiological/biological role are possible. Recently BZD-receptor binding activity equivalent to surprisingly high levels of more than 900 ng/ml was found in cerebrospinal fluid of patients with advanced hepatic encephalopathy. As long as the structure of this binding activity has not been elucidated no firm conclusions can be drawn from these findings. If pertinent analytical problems (e.g. drugfree biological material; exact quantification by internal standard techniques) are solved and if the source(s) of BZD are established it might be possible to answer also the critical question whether “endogenous” or “natural” BZD play any (in-) direct role in the regulation of CNS activity. Many drugs are of a natural plant origin (e.g. alkaloids). More recently it has been elucidated that morphine and codeine are present in different mammalian tissues (1) and the skeleton of morphine can be synthesized by the rat liver (2). Similarly, benzodiazepines (BZD) represent widely used drugs with a complex chemical structure. Their pharmacological effects are mediated by specific BZD-GABAA-chloride channel receptors (3) whose molecular structures have recently been described (4, 5). Since the discovery of these receptors in 1977 there is an ongoing discussion whether endogenous ligands exist and various putative candidates have been hypothesized (6, 7). The latest contributors to this long list are surprisingly BZD. Therefore it is the aim of this minireview to discuss the natural occurence of these “endogenous” BZD.

note the 1 ng/g and for DD from 0.01 to 0.5 ng/g

 

[vecktor]

if someone had the research grant the question could be answered. The most direct way would be to look for 14C isotope ratios because the phenyl ring in synthetic benzodiazepines will have been made from crude oil whereas plant produced materials will have been produced from photosynthetic products (probably within the last growing season). Thanks to atmospheric nuclear tests there is a nice spike in 14C in all plant matter from the 1950's onwards. If the ratio suggests recent synthesis from a photosynthesis product then that is very strong evidence of it being natural.

I'm pretty sure though this is contamination, as chlorine containing compounds are somewhat rare in nature and there are a whole lot of these benzodiazepines being found all of which are drug metabolites. wierder things have happened for example the discovery that morphine is naturally synthesized in humans.

 

[Jamshyd]

A few years back there was a study that showed that Potato contained trace amounts of Bromazepam and other benzos. I will try to dig it out if no one beats me to it...

 

[Swerlz]

Planta Med. 2008 Apr;74(5):491-6.
Potato (Solanum tuberosum) juice exerts an anticonvulsant effect in mice through binding to GABA receptors.

Muceniece R, Saleniece K, Krigere L, Rumaks J, Dzirkale Z, Mezhapuke R, Kviesis J, Mekss P, Klusa V, Schiöth HB, Dambrova M.

University of Latvia, Faculty of Medicine, Sarlotes Str. 1a, Riga, Latvia. [email protected]

Abstract

Naturally occurring benzodiazepines have been identified in regular food such as wheat and potato, but there is still no evidence that potato extracts can affect CNS responses in vivo. Here we found that undiluted potato juice and potato juice diluted with saline 1 : 2 administered 10 min intracisternally ( I. C.) and 30 min per os before bicuculline exerted significant anticonvulsant activity in the bicuculline-induced seizure threshold test in mice. In vitro, potato juice from different harvests at dilution series from 10 % to 0.000001 %, diluted 100,000-fold, displaced 50 % of gamma-aminobutyric acid (GABA) receptor ligand [ (3)H]GABA and diluted 40-fold displaced 50 % of [(3)H]flunitrazepam from binding sites in mice forebrain membranes. The low content of diazepam (0.04 +/- 0.01 mg/kg) determined by HPLC and mass spectrometry in the potato extracts could not sustain the anticonvulsant activity of potato juice in vivo; therefore we hypothesized that potato juice might contain GABA (A) receptor GABA-site active compounds. The findings of this study suggest that potato juice as well as potato taken as food may have the capacity of influencing brain GABA-ergic activity.

source

 

[Swerlz]

Biochem Biophys Res Commun. 1988 Dec 30;157(3):1436-43.
Increase of natural benzodiazepines in wheat and potato during germination.

Wildmann J.

Zentrum Biochemie, Georg-August-Universität Göttingen, West Germany.

Abstract

Aqueous acid extracts of wheat grains and potato exhibit after HPLC separation a series of compounds that are able to inhibit the binding of benzodiazepines to benzodiazepine receptors of rat brain membranes. In wheat one of the inhibiting compounds was shown to be identical to diazepam by means of HPLC characterization and gas chromatography combined with mass spectrometry. In potato one of the most prominent components in terms of binding inhibiting activity was identified as lormetazepam. In wheat and potato germination increases total inhibiting activity of the whole plant extracts as well as the content of the benzodiazepines approximately by factor five. Because uptake of benzodiazepines from the surrounding was excluded these findings indicate the biosynthesis of the benzodiazepines diazepam and lormetazepam by the plants investigated.

source

 

[(zonk)]

i doubt its pollution, an easier way would be to grow these in a lab where no pollutants could possible effect them and see the concentrations

 

[Phener]

The wonders of HPLC! Not buying it one bit!

Ok so you are an academic:

• Own an HPLC machine
• Haven't published for a while
• Have students under you who need to learn HPLC

Because uptake of benzodiazepines from the surrounding was excluded

• Details, I haven't read the study but I highly doubt they grew potatoes from scratch in soil which had been thoroughly screened for any benzodiazepine
• More than likely this would involve a simple *take sample of surrounding soil* compare and contrast any trace benzodiazepine levels

Interesting how only manmade (or metabolites of) benzodiazepines have been discovered and at TRACE levels.

Don't get me wrong there is a BZD receptor in the brain so highly likely there are endogenous agonists BUT crudely using an HPLC machine to test crop samples (finding only nanogram levels of know benzos) is hardly cutting edge proof. Equa

Abstract

Naturally occurring benzodiazepines have been identified in regular food such as wheat and potato, but there is still no evidence that potato extracts can affect CNS responses in vivo. Here we found that undiluted potato juice and potato juice diluted with saline 1 : 2 administered 10 min intracisternally ( I. C.) and 30 min per os before bicuculline exerted significant anticonvulsant activity in the bicuculline-induced seizure threshold test in mice. In vitro, potato juice from different harvests at dilution series from 10 % to 0.000001 %, diluted 100,000-fold, displaced 50 % of gamma-aminobutyric acid (GABA) receptor ligand [ (3)H]GABA and diluted 40-fold displaced 50 % of [(3)H]flunitrazepam from binding sites in mice forebrain membranes. The low content of diazepam (0.04 +/- 0.01 mg/kg) determined by HPLC and mass spectrometry in the potato extracts could not sustain the anticonvulsant activity of potato juice in vivo; therefore we hypothesized that potato juice might contain GABA (A) receptor GABA-site active compounds. The findings of this study suggest that potato juice as well as potato taken as food may have the capacity of influencing brain GABA-ergic activity.

^this study is hinting at OTHER gabaergic compounds but again it's ONE study and if you read it fully I bet you the science will be VERY speculative. Note 0.01mg/kg = 10nanograms/g

Academics need to regularly publish. People need to remember this.

i doubt its pollution, an easier way would be to grow these in a lab where no pollutants could possible effect them and see the concentrations

yes but did they bother to do this? ...I'm guessing not!

 

[wungchow]

There are a number of plants/organisms that can synthesize the 1,4-benzodiazepine core, but apparently bacteria are the best.

This is the biosynthetic pathway proposed for formation of diazepam-like BZDs.

They claim that halogens can be further introduced by bacterial haloperoxidase enzymes.

(source)

 

[wungchow]

Another interesting thing to note is diazepam-like BZDs have been detected postmortem in the brains of a subjects who died before 1959 (clinical introduction of benzodiazepines). (source)

 

[wungchow]

Another plausible biosynthetic route involves formation of a cyclic dipeptide with anthranilic acid and glycine...

 

[(zonk)]

So if these are produced in nature(which dont think is farfetched at all)does this mean the patents(for the ones discovrd)are now void. Anyone wanna market some "herbal"benzo "extacts" $$$

 

[Phener]

So if these are produced in nature(which dont think is farfetched at all)does this mean the patents(for the ones discovrd)are now void. Anyone wanna market some "herbal"benzo "extacts" $$$

that would be one expensive diazepam tablet!
20,000,000kg of potato extract = 5mg
(even 2000kg of highly bread plant would be one expensive tablet)
agreed you could probably get genentically modified bacteria on the job but then they have said the same for psycadelics like LSD (not going to happen clandenstinely, not going to happen by big pharma)


Personal opinion:
Academics have been for a while DESPERATE to find benzodiazepines in mamallian brains as proof of endogenous BZD receptor agonists. Most of the publications are speculative on their biosynthetic pathways and all their HPLC methodology are open to flaws (or extremely biased science for a quick publication to add to ones neuro CV of publications).

I would predict if an endogenous agonist exists that it is probably a peptide or rather more complicated molecule than a simple known BZD. Think Anandamide, 2-Arachidonoylglycerol (2-AG) (CB receptors agonists), endomorphins (opioid receptor peptides)

 

[vecktor]

.

I would predict if an endogenous agonist exists that it is probably a peptide or rather more complicated molecule than a simple known BZD. Think Anandamide, 2-Arachidonoylglycerol (2-AG) (CB receptors agonists), endomorphins (opioid receptor peptides)

B carbolines are found in vivo in much higher concentrations than these 'natural' benzodiazepines and some of them are very decent ligands I think that is where people need to look for the endogenous BZD ligand.

 

[(zonk)]

Well i wasn't actually suggesting performing an extraction, hence the ""s on "extraction". I was thinkn synthesizing them in a lab and calling it "extract". No one could ever proove otherwise

 

[MagickalKat777]

if someone had the research grant the question could be answered. The most direct way would be to look for 14C isotope ratios because the phenyl ring in synthetic benzodiazepines will have been made from crude oil whereas plant produced materials will have been produced from photosynthetic products (probably within the last growing season). Thanks to atmospheric nuclear tests there is a nice spike in 14C in all plant matter from the 1950's onwards. If the ratio suggests recent synthesis from a photosynthesis product then that is very strong evidence of it being natural.

Well i wasn't actually suggesting performing an extraction, hence the ""s on "extraction". I was thinkn synthesizing them in a lab and calling it "extract". No one could ever proove otherwise

See above quote.

Anyway, I am on the skeptic side as well. I highly doubt there will ever be a natural benzodiazepine. The only thing that comes close is valerenic acid and the data on it is few and far between - they believe it binds to GABAA BZD receptors but haven't really researched agonist/antagonist properties - and the dose required for effects is ridiculous.

 

[Timothy Leary]

The highest report I have seen here is 1 ng/g, that would mean that even if this stuff had the per gram potency of LSD, you would have to use 20 kilograms to get a threshold dose. This is.basically useless, the only importance is for the legal implications for big pharmaceutical companies. I bet if you factor in environmental stability of different Benzos and how much is excreted by humans without breaking down extensively that the ratios of which Benzo it is changes in line with the description rates of certain ones. maybe in 1970 it was vallium more, the 80s lorezapam, 90s to now, xanax, I bet it varies by country as well.

 

[hamhurricane]

^^^
Right, but why just diazepam/nordiazepam then especially when one of these studies was conducted in 1989? I'm aware benzos are most prescribed class of psychoactive medicine (or at least were) but surely you would see all kinds of other things as well if it was simply an environmental pollutant. Pharmaceutical pollution surveys have detected dozens of drugs in drinking water internationally e.g. Bezafibrate, Bleomycin, Clofibric acid, Carbamazepine, Diclofenac, Gemfibrozil, Ibuprofen, Phenazone, Propylphenazone and Tylosin

Pharmaceuticals: a threat to drinking water?
Oliver A. Jones, John N. Lester and Nick Voulvoulis

 

[Timothy Leary]

Perhaps starch rich plants (vegetables, grain, corn), like the ones they tested like to concentrate them, I suppose the isolation techniques could have filtered a lot out, I mean you have to come up with a pretty good extraction to get all that plant matter and natural secondly metabolites.