Chemical Analysis of Galbulimima Bark

[Amontaiyagala]

The genus Galbulimima (Himantandraceae) consists of large rainforest trees distributed in Australia (North Queensland), Papua New Guinea, and the Moluccas. Depending on the botanical authority 1-4 different species were acknowledged, but Australian G. baccata and New Guinean G. belgraveana are most often accepted. Previous reports have indicated that in Papua New Guinea the bark of G. belgraveana was ingested by specialized diviners to receive dreams of prediction (Hamilton 1960, Trans. PNG Sci. Soc. 1, 16-18). Self experiments have previously been described here: http://www.shaman-au...showtopic=19895

Although more than 28 alkaloids have been isolated from the bark so far (Brown et al. (1956) Aust. J. Chem. 9, 283-287; Binns et al. (1965) Aust. J. Chem. 18, 569-573) the active principle has remained unidentified.

I became interested in Galbulimima belgraveana in 1994 while traveling in Papua Niugini. During extensive fieldwork in remote areas of the Eastern Highlands in 1999 and 2004 I managed to collect several samples under the supervision of practicing shamans who also introduced me into eating the bark (it can be VERY active).

Fore shaman holding Agara bark (G. belgraveana) taken from a tree considered to be inactive because of its location in a recently cleared garden site.

What might be particularly interesting with respect to the (ethno)botany of Galbulimima is that shamans differentiate between two kinds of trees, described as “man” & “meri” (male and female) although the trees are clearly monoecious. Furthermore, shamans employ a rigorous selection procedure to identify trees suitable for the preparation of the bark drug which takes morphological, ecological as well as supernatural considerations into account. Only trees from the primary rainforest at the top of mountain slopes are employed and those in areas of “ples masalai” (spirit places) are considered to be particularly powerful. It is generally assumed that Galbulimima trees exposed to frequent human contact loose their power because they are abandoned by the tree spirit who is believed to reside in them.

Most interestingly, shamans selection procedures seem to be paralleled by an extreme variability in alkaloid composition observed during chemical analysis:

“Whether the genus consists of one, two, three, or four species the variability of alkaloid content with respect both to total amount and to individual alkaloids is striking and possibly unique. Of the eleven bark samples from North Queensland and the seven from New Guinea that have been worked up for alkaloids no two have been alike. Even samples collected in the same area and at approximately the same season have been quite dissimilar. The variability thus revealed by actual isolation of the alkaloids has been confirmed by a paper-chromatographic survey of the alkaloids of an extensive series of samples. The yield of total alkaloids varied from a trace to about 0.5%. The major and commonest alkaloids in bark from both sources were himbacine and himandridine. Himbadine and himgaline, which are structurally related, were also major alkaloids in bark from North Queensland but were not found in material from New Guinea, except in one instance when a trace of himgaline was isolated. Himbosine was usually present in small amounts in all samples but all the other alkaloids occurred irregularly and should be regarded as minor. Some have been isolated only once.” Ritchie & Taylor (1967) in: The Alkaloids Vol.9, RHF Manske (Ed.), 529-543.

Major alkaloids from New Guinean G. belgraveana and Australian G. baccata.

Recently, I regained the opportunity to perform chemical analyses using state of the art analytical techniques (NMR and HPLC-MS) and have started to analyze my samples that had been stored away for many years at -20 °C.

For isolation of crude Galbulimima bark alkaloids I employed the following extraction procedure: 1 g of dried bark was powdered and extracted with 3 x 10 ml methanol for 24 h at room temperature. The filtered extract was concentrated to dryness in vacuum and the residue taken up in 1 ml 5% aqueous hydrochloric acid and extracted with 3 x 1 ml diethyl ether. The aqueous phase was basified with 1 ml 20% aqueous ammonia solution and extracted with 3 x 1 ml chloroform. The organic phase was dried over sodium sulfate, filtered and concentrated to dryness in vacuum. The residue was taken up in 600 µl chloroform-d₁ and analyzed by NMR spectroscopy.

The 500 MHz proton NMR spectrum suggested that the crude alkaloid extract consists of almost pure Himgravine (approx. >90%). Isolated amounts exceed 10 mg/g of dried bark, which means that alkaloid content in this sample is at least twice as high as those reported in the literature.

¹H NMR spectrum (500 MHz, CDCl₃) of crude G. belgraveana alkaloids shows Himgravine as the major component.

However, as no NMR data of authentic Himgravine have ever been published (apart from a very low resolution ¹H NMR in Abraham & Bernstein (1961) Aust. J. Chem. 14, 64-68), I have also acquired a H,H-COSY spectrum to confirm the tentative identification of Himgravine. Again, the data are in excellent agreement with the proposed identification of Himgravine. However, I intend to acquire additional NMR spectra (¹³C NMR, HMQC, HMBC, and NOESY) to unambiguously establish the structure assignment and will sure run HPLC-MS to get an idea about the minor components. In addition, I will continue to analyze my various other G. belgraveana samples (man & meri, active & inactive) and might also try to isolate some of the alkaloids in pure form.

H,H-COSY spectrum (500 MHz, CDCl₃) of crude G. belgraveana alkaloids shows Himgravine as the major component.

Please keep in mind that the analyzed sample is derived from a tree that was not used for divination purposes, so I do not claim that Himgravine is an active component. However, I do hope that analysis of my other samples (some of which are surely active) will enable identification of the active principle(s).

What I’m now interested in is to get access to additional samples of authentic Galbulimima bark from Papua Niugini and Australia. The idea is to analyze active as well as inactive specimen in order to find candidate compounds for the observed activity and thus to bypass bioassays with all the different isolated components, some of which might in fact be harmful (especially if taken on their own). Consequently, samples which have already been bioassayed with positive as well as negative results would of course be most helpful. Amounts as low as 1 g of dry bark could be sufficient to identify the major component(s). Crude alkaloid mixtures could be send back if requested. Please be advised that I will NOT perform extractions of large bark samples for you – my intention is chemical analysis only. I’m also not interested in analyzing anything else apart from Galbulimima (no need to ask, sorry) and I will perform microscopic analysis of bark samples prior to any extraction – samples other than Galbulimima would be discarded.

For those of you who feel inspired to visit Papua Niugini on their own in order to acquire G. belgraveana, I would like to give you the following advice:

• Learning Tok Pisin (Pidgin, Neo-Melanesian) is absolutely essential because people capable of speaking English do not know anything about traditional customs and plant use (translators are not suitable).
  
• Keep in mind that you are most likely to fall prey to con-men who will try to deprive you of your personal belongings.
  
• Don’t expect to obtain any (authentic) samples during your first visit. Plan at least several weeks living with the indigenous people before a reliable level of trust has been established.
  
• Sampling of any tree requires the consent of village leaders and landowners.
  
• Keep in mind that Galbulimima is used to divine the cause of sickness, which is most often considered to be sorcery. Your interest in knowledge of sorcery and counter sorcery makes you vulnerable to serious sorcery accusations and might severely endanger your safety.
  
• Comply with quarantine regulations.
  
• Ingestion of any plant material might pose serious health risks including permanent psychological or physical damage, or even death.
  
• Please stay safe.
  

 

Edited September 1, 2012 by Amontaiyagala

[CβL]

I get the feeling that Himgravine and Himbacine are likely to be the main two components, based on how they look (nice and flexible, vaguely similar to xanthines). ^^

But thanks very much for your rigorous treatment of the subject, it's very much appreciated, and I only wish I could help.

[tarenna]

Brilliant work. I have a cultivated specimen that is about 4m tall - ex Atherton Tableland. No idea of activity or alkaloid content. If interested in a bark specimen please pm me.

Good luck

[mindperformer]

Very resourceful reports!

Himbacine, an alkaloid, is a strong competitive M2-muscarinic antagonist and therefore against Alzheimer.

Atropine, Scopolamine, Diphenhydramine, Speciogynine, Convolvine and many other compounds are also antagonistic on this receptor.

Many plants against Alzheimer (like Galantamine from the Red Spider Lily) induce dreams, this needs more investigation.

Edited September 1, 2012 by mindperformer

[CβL]

Actually, after staring at Himbacine for a while (lol), I noticed it's similar in structure to Pukateine (takes a while to see it)

Pukateine (rotate it ccw 90 degrees).

Himbacine

If we could take Himbacine's "tail", and bond it backwards to the main rings to make the structure more rigid, then maybe it would have more of an effect on D2 receptors (as Pukateine does).

I also wonder if it's just coincidence that one of my higher-dosage bioassays with Pukateine gave me a rather powerful dissociative experience.

[mindperformer]

very interesting, i thought so too, that there is some similarity with antipsychotics...

Did you experience with pure Pukateine? What was your dosage?

Was it sedative-dissoziative or rather stimulating-dissoziative?

I've bee searching for the tree which contains it (Laurelia novae-zelandiae) when I was in New Zealand but there wasn't enough time for the northern part of the north-island. Now there are sellers of the bark and I am thinking on ordering some of it. A Maori in NZ told me they use it against pain of all kind.

Salvinorin A is also a very potent (partial) agonist on the D2-receptor, like Pukateine, and this could possibly explain the dissoziative action (Salvinorin A is additionally a less strong kappa-opioidreceptor agonist).

Edited September 2, 2012 by mindperformer

[Amontaiyagala]

Actually, after staring at Himbacine for a while (lol), I noticed it's similar in structure to Pukateine (takes a while to see it)

I would not take these chemical structure drawings too serious if similarity is concerned - at least not if it is about pharmacological similarity. In 3D Pukateine is pretty flat, whereas Himbacine is not. Furthermore, taking a closer look at the molecular orbitals of Pukateine and Himbacine (both as the ammonium salt likely to form under physiological pH) shows that their Highest Occupied Molecular Orbitals (HOMOs) have nothing in common.

So far Himbacine has been tested for binding at muscarinic receptors only, where it shows some selectivity for M2 and M4 receptors. There is, however, some evidence that Himbacine might affect dopamine release via indirect pathways.

[CβL]

Thanks for that detailed answer.

Without being very knowledgeable with Structure-Activity Relationships, I don't think the case is settled for their relationships being dissimilar.

Secondly, there's other alkaloids from Pukatea that could perhaps be related (Laureline is also

a breakdown product of Pukateine). I'm meant to be writing a report so don't have time to check it out, but here's a paper with a list of alkaloids from the species:

Laurelia novae-zelandiae A. Cunn.Obovanine (25), oxoputerine, (-)-romneı́ne (8), pukateine (16), laureline (27), liriodenine (44), estepharine (50), pukateine-N-oxide, pukatein-methylether, isolaureline, laurolitsine (21), mecambroline (23), boldine (12), isoboldine (19), roemerine (17), oxolaureline, oxoputerine (38), lauterine, coridine (13), (-)-laurepukine (32), laurepukine N-oxide, stepharine (49)

Some damn interesting Lignoids from the family:

http://www.sciencedirect.com/science/article/pii/S0378874198002335

[mindperformer]

very interesting!

all in all boldine-like alkaloids.

Many of the constituents are dopaminergic:

Dopamine- Receptors: D₁, D₂, D₃, D₄, D₅

Endogene ligands: Dopamine

Psychoactive substances:

Agonists:

(D2-Agonism inhibits NMDA-R., therefore hallucinogenic)

PCP (partial), Apomorphine,

Ketamine (partial), LSD,

Cestrum parqui- Extract,

Asarone, Atherosperminine,

Agroclavine, 2-Aminoindan,

R-Aporphine, Bromocriptine,

Pukateine, Vitexlactam A,

Salsolinol, Salvinorin A (D2),

Cabergolin (aphrodising through D2),

Mescaline,

Antagonisten: Haloperidol, Paeonol,

Tetrahydropalmatine

(enhances D in other areas),

L-Stepholidine, Boldine,

Glaucin, Methcathinone,

Nuciferin, Rotundin,

Tetrahydroberberine,

Hederagenine, Clozapine,

Prothipendyl (Dominal),

Tetrahydro-columbamine,

water-fraction from

Acorus gramineus, d-

Coclaurine,

Inverse Agonists: Flunitrazepam,

Edited September 8, 2012 by mindperformer

[mindperformer]

The parade example LSD also works on the D2-receptor, so this could be responsible for some of its hallucinogenic properties, especially the dissociative part:

http://www.ncbi.nlm.nih.gov/pubmed/9698051

[CβL]

Okay, I can't not explain why it's so awesome...

But look at the molecule 103 - Yatein. It's very similar to two certain, well-revered, phenethyamines joined together. I'm not at all suggesting it would as a combination act as such (it 99.99999% won't), but maybe with a cleave in the right place, it could become a something useful for further processing... I'm not at all suggesting that yatein is the ideal molecule for such a cleave, but rather that certain more common lignans may possess more amenable structures that when cleaved result in rare molecules.

/\

This related bad-boy is used to treat genital warts, and has anti-cancer properties. Hehehehehe

[tempus]

Himbacine is a selective M2/M4 antagonist. This means that it is able to significantly increase locomotion without the cognitive effects mediated by antagonism of the M1 receptor by other muscarinic acetylcholine antagonists such as Atropine, Scopolamine and Diphenhydramine. Half life is also fairly long with an effective duration between 24 and 48 hours.

In my opinion Galbulimima is one of the best medical herbs against fatigue-related health problems.