Her benyttes photosensitizeren TPPS2a, husker ikke i farten om PCIB har rettighetene til denne. PCIBs patenterte photosensitizer, fimaporfin, er TPCS2a.
Ser PCIB er nevnt en gang i artikkelen, ser ut som om de har lånt ut sin patenterte LumiSource lampe som brukes for å belyse.
Det blir mye prat på småprat så jeg legger denne ut her. Er dere informert om denne?
Photo-triggered theranostic
Om vi ser under så er det aktivitet mellom Multimmune og PCIB
Fantastiske resultater! (ikke sett før selv, de er fra 2010)
http://europepmc.org/article/MED/32053965#sec5-cancers-12-00417
In conclusion, this is the first report demonstrating cytosolic delivery of a small-moleculeinhibitor by PCI. Sunitinib-PCI was found highly promising in vitro when utilizing the “light first” protocol and was also indicated to increase treatment-induced necrosis in vivo. The overall treatment response in our animal models was, however, less than expected which indicate mechanisms in the tumor stroma to attenuate an overall treatment response. Particularly in CT26.WT tumor-bearing thymic animals where an antagonistic effect on infiltrating T-cells was observed. Hence, our work indicates PCI to potentiate sunitinib cytotoxicity although adjuvant therapy aimed at the tumor stroma should be evaluated to improve the therapeutic efficacy.
Fra may 2015.
Photosensitizer and Light Pave the Way for Cytosolic Targeting and Generation of Cytosolic CD8 T Cells Using PLGA Vaccine Particles
Feb.2020
Sitat
´´These data suggest that the drug-loaded TPC−CS NPs have a potential in combinatory anticancer therapy and as contrast agents.´´
Kan jo kanskje være lurt å skrive litt mer og utdype hvorfor du mener hva det mest relevante for PCI Biotech er?
Takk. Ok, trodde fundamental tråd ikke ga rom for det. Ville bare vise mangfold av muligheter PCIB har.
Har gått meg litt hus forbi hvor bra dette er sjølv med det godt forklaret innlegget til Investor.
Ein øker overlevelsen fra ein plass mellom 11 til 12 mnd til ein plass mellom 17 til 23 mnd. PW sine egne ord i podcast 117. (15 min inn)
Hørt igjen på podcast 128 fra 50 min til 51 min og endelig skjønner eg også hvorfor denne patenten var så viktig. “der vi ser veldig veldig sterke synergier” “1 pluss 1 blir ikkje 2 men 7” og hør på den latteren når Elisabeth seier “det er bra”. Får ein følelse av at PW har lyst å sei det er jævla bra.
Jeg må krype til korset her. Jeg hadde ikke fått med meg dette fra extension cohort.
Dette er sykt sterkt.
Edit:
Dette highligheter hvorfor vi trenger fundamentaltråden, mye større sjanse for at det faktisk løftes frem relevant informasjon.
Jeg skrev det på chatten tidligere i dag, forumet fortjener ikke @Investor
Er litt som å ha @Rodjer i PHO, bare spar opp gull etter gull
Et ytterligere punkt jeg kan føye til i min tro på AZD8601 som medisinen FimaNAc er prøvd ut på, er at selve koblingen mellom AstraZeneca og Moderna er fundert på AZD8601 og forskeren Kenneth R. Chien som pr. i dag er ansatt på Karolinska og jobber for begge de to selskapene:
I 2010 grunnlegges selskapet Moderna av seks forskere og et venturefond (https://www.flagshippioneering.com/ som også ut over Moderna har en imponerende track record!). Kenneth R Chien er en av disse seks, men han fortsetter i sin jobb som professor på Harvard.
I 2012 blir Chien ansatt som professor i kardiovaskulær forskning vid Karolinska for å lede Integrated Cardio Metabolic Centre, ICMC. I følge arbeidskontrakten hans med KI, skal han bruke 20% av tiden sin som rådgiver for AstraZeneca: https://drive.google.com/file/d/0By2HqPi4t2RbS2pVdE1pbDFEQjV1QUJfcGZvRnR4R196eE9B/view og https://drive.google.com/file/d/0By2HqPi4t2RbTkUyZXFDWDVtT2l6NUl5LW9BUUVoQmVYUDEw/view
I 2013 investerer AstraZeneca USD240+180 millioner i Moderna. I følge denne kilden, som et direkte resultat av AstraZeneca sin begeistring for VEGF mRNA forsøkene: https://www.sciencemag.org/news/2017/02/mysterious-2-billion-biotech-revealing-secrets-behind-its-new-drugs-and-vaccines
I 2015 donerer Ming Wai Lau, (sønn av Joseph Lau, den fjerde rikeste personen i Hong Kong) 450 millioner(!) SEK til KI, som var øremerket Chen sin forskning, samt å få hjelp til å bygge opp Ming Wai Lau Center for Regenerative Medicine Hong Kong. Denne saken har skapt masse rabalder Sverige.
ICMC gjør de kommende årene oppdragsforskning for både AstraZeneca og Moderna. Det meste som ICMC gjør er hemmelig, men av de artiklene som blir publisert, er det tydelig at de hovedsaklig jobber med VEGF mRNA. Veldig mange av dem er i samarbeid med forskere fra det nyopprettede instituttet i Hong Kong.
Her er et utvalg:
https://www.nature.com/articles/s41467-019-08852-4
Mye info, men det store poenget her er at en av grunnleggerene av Moderna og oppfinneren av AZD8601 sitter i Sverige og jobber tett med AstraZeneca, og vil naturligvis ha et veldig høyt ønske om at hans medisin til slutt når markedet, og som det står i en av han artikler (fra i fjor sommer!): “novel delivery approaches of modRNA are needed to improve therapeutic efficacy in the diseased setting”.
Jeg blir bare mer og mer overbevist om at AZD8601 ble forsøkt levert med TPCS2A og laser før jul, og jeg er sterk i troen på at de også har fått det til. Og om det stemmer, så vil både Moderna og AstraZeneca ønske å sikre seg denne teknologien - ikke bare lisensiere den!
Må bare slutte meg til @sjog.
Om du har rett eller ikke; dette er jobbing og graving på høyt nivå. Gålr dine antakelser fullt inn, er det bare å spenne sikkerhetsbeltene hardt til de nærmeste ukene.
Krysser fingrene for at ditt sherlock arbeid stemmer.
Långt inlägg…
Något som inte tas upp lika mycket är CRISPR, gensaxen som många tror kommer revolusionera medicin. Svårigheten ligger som vanligt i leverering 
Från Thermi Fishers hemsida, de producerar, Lipofectamine som används i försök inom in vivo försök med CRISPR.
Transfection is the process by which CRISPR-Cas9 DNA, mRNA and protein systems are introduced into eukaryotic cells. Techniques vary widely and include lipid nanoparticle–mediated transfection, viral delivery, and physical methods such as electroporation. Our Invitrogen Lipofectamine family of reagents paired with the Neon Transfection System offer complete delivery solutions to address your genome editing needs. We have optimized protocols for each product option to help you achieve high cleavage efficiency and the ease of delivery you expect.
Om man sedan går till forskningsartiklar som finns låter det inte lika övertygande om lipofectamine.
Delivery Aspects of CRISPR/Cas for in Vivo Genome Editing. Danny Wilbie et al. Acc Chem Res. 2019.
In this Account, we focus on the delivery aspects of CRISPR/Cas for therapeutic applications in vivo. Safe and effective delivery of the CRISPR/Cas components into the nucleus of affected cells is essential for therapeutic gene editing. These components can be delivered in several formats, such as pDNA, viral vectors, or ribonuclear complexes. In the ideal case, the delivery system should address the current limitations of CRISPR gene editing, which are (1) lack of targeting specific tissues or cells, (2) the inability to enter cells, (3) activation of the immune system, and (4) off-target events.
fimaNac gör det möjligt att 1: aktivera lokalt, 2: ökar möjligheten att komma in i cellerna/cytoplasman och 3: därmed få önskad effekt där man önskar och samtidigt minska oönskade effekter på andra platser.
To circumvent most of these problems, initial therapeutic applications of CRISPR/Cas were performed on cells ex vivo via classical methods (e.g., microinjection or electroporation) and novel methods (e.g., TRIAMF and iTOP). Ideal candidates for such methods are, for example, hematopoietic cells, but not all tissue types are suited for ex vivo manipulation. For direct in vivo application, however, delivery systems are needed that can target the CRISPR/Cas components to specific tissues or cells in the human body, without causing immune activation or causing high frequencies of off-target effects . Viral systems have been used as a first resort to transduce cells in vivo. These systems suffer from problems related to packaging constraints, immunogenicity, and longevity of Cas expression, which favors off-target events. Viral vectors are as such not the best choice for direct in vivo delivery of CRISPR/Cas. Synthetic vectors can deliver nucleic acids as well, without the innate disadvantages of viral vectors. They can be classed into lipid, polymeric, and inorganic particles, all of which have been reported in the literature. The advantage of synthetic systems is that they can deliver the CRISPR/Cas system also as a preformed ribonucleoprotein complex. The transient nature of this approach favors low frequencies of off-target events and minimizes the window of immune activation.
Moreover, from a pharmaceutical perspective, synthetic delivery systems are much easier to scale up for clinical use compared to viral vectors and can be chemically functionalized with ligands to obtain target cell specificity. The first preclinical results with lipid nanoparticles delivering CRISPR/Cas either as mRNA or ribonucleoproteins are very promising. The goal is translating these CRISPR/Cas therapeutics to a clinical setting as well. Taken together, these current trends seem to favor the use of sgRNA/Cas ribonucleoprotein complexes delivered in vivo by synthetic particles.
Troligtvis kan fimaNac finna en plats även här.
Delivering CRISPR: A Review of the Challenges and Approaches
Christopher A Lino et al. Drug Deliv. 2018 Nov.
Abstract
Gene therapy has long held promise to correct a variety of human diseases and defects. Discovery of the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR), the mechanism of the CRISPR-based prokaryotic adaptive immune system (CRISPR-associated system, Cas), and its repurposing into a potent gene editing tool has revolutionized the field of molecular biology and generated excitement for new and improved gene therapies. Additionally, the simplicity and flexibility of the CRISPR/Cas9 site-specific nuclease system has led to its widespread use in many biological research areas including development of model cell lines, discovering mechanisms of disease, identifying disease targets, development of transgene animals and plants, and transcriptional modulation. In this review, we present the brief history and basic mechanisms of the CRISPR/Cas9 system and its predecessors (ZFNs and TALENs), lessons learned from past human gene therapy efforts, and recent modifications of CRISPR/Cas9 to provide functions beyond gene editing.
We introduce several factors that influence CRISPR/Cas9 efficacy which must be addressed before effective in vivo human gene therapy can be realized.
The focus then turns to the most difficult barrier to potential in vivo use of CRISPR/Cas9, delivery.
We detail the various cargos and delivery vehicles reported for CRISPR/Cas9, including physical delivery methods (e.g. microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles), and discuss their relative merits. We also examine several technologies that, while not currently reported for CRISPR/Cas9 delivery, appear to have promise in this field. The therapeutic potential of CRISPR/Cas9 is vast and will only increase as the technology and its delivery improves
.
Flera dödsfall i tidiga försök med genterapi:
It is therefore critical that gene therapy technologies allow for highly specific editing of the genome to reduce the risk of undesired mutagenesis, and that the delivery vehicle allows for safe and efficient transport to the target.
.
fimaNac kan troligen hjälpa till att få önskad effekt med ökad precision, dvs ändra genomet lokalt och inte på oönskade platser i kroppen.
Careful consideration and development of both the gene editing tool and the delivery mechanism will be required if the full potential of therapeutic gene editing is to be realized.
Crispr och lipid nanoparticles.
There are substantial drawbacks for delivery of CRISPR/Cas9 components via lipid nanoparticle. First, there are both external and internal barriers that must be considered. Once the nanoparticle has passed through the surface of the cell, it is typically encased within an endosome. Encased contents can very rapidly be directed by the cell into the lysosomal pathway, causing the degradation of all lysosome contents. Therefore, the cargo must escape the endosome. Also, if the Cas9:sgRNA complex can escape the endosome, it must also translocate to the nucleus, which can also be a potential point of failure.
Because of this, it is rare to see particularly high efficacies when delivery CRISPR/Cas9 components via lipid nanoparticles. While Wang et al. (2016) could achieve ∼70% in vitro modification efficiency in cells (see Figure 7(A)), that only came after an intense screen to determine the most optimal lipids with which to construct their liposomes for their system. Finally, lipid nanoparticles are like virus particles in that the nature and size of the cargo, along with the target cell type, highly affect transfection efficiency and the types of lipids that are appropriate or useful in the system.
Because of their lack of viral components, there will always be interest in improving lipid nanoparticles to deliver CRISPR/Cas9 components. This improvement process can come through the screening of better lipid carriers, as above; better decorations on the liposome surface to help target particles to specific cells or tissues, avoid immune system detection, and facilitate endosomal escape; and improved packaging of CRISPR/Cas9 components, increasing the odds of some subset of packaged molecules to be appropriately delivered.
Other common polymeric vectors for DNA delivery are polyethenimine (PEI) and poly(l-lysine) (PLL). Branched PEI have high charge density, facilitating efficient plasmid DNA packing, and pH-buffering ability which enables escape from endosomes. However, branched PEI is cytotoxic. Therefore, a balance between the desirable properties of branched PEI and the less toxic linear PEI must be struck for effective transfection.
Återigen svår balans mellan effekt och toxicitet.
Refererar till PCIB:s presentation om fimaNac: Enhancement by fimaNac is best under conditions favourable for vehicle safety.
Low concentration of vehicle/nucleic acid complex.
Låg koncentration av den delen som har toxiska egenskaper ger bäst effekt, och det är då den fungerar som bäst. Vanligtvis måste man ligga på gränsen till toxiska reaktioner för att få önskad effekt, här är det tvärt om. Helt otroligt.
Citat från samma presentation som ovan:
fimaNac is especially advantageous in vivo :
- difficult to achieve a high concentration of vehicle/nucleic acid complex in target cells.
- Toxicity may limit the amount of vehicle used.
Det återkommer hela tiden inom genterapi oavsett typ: toxiciteten begränsar. Svårigheten är att leverera!
Mycket tyder på att det nu är löst för i alla fall vissa indikationer.
Över till AZ och CRISPR
Thursday, 29 January 2015
AstraZeneca today announced four research collaborations aimed at harnessing the power of CRISPR, a pioneering genome-editing technique, across its entire discovery platform in the company’s key therapeutic areas. The technology will allow AstraZeneca to identify and validate new drug targets in preclinical models that closely resemble human disease. AstraZeneca will share cell lines and compounds with its partners and work with them to publish findings of its application of CRISPR technology in peer-reviewed journals, contributing to broader scientific progress in the field. The collaborations complement AstraZeneca’s in-house CRISPR programme and will build on the company’s ‘open innovation’ approach to research and development.
CRISPR (clustered regularly interspaced short palindromic repeats) is a genome-editing tool, which allows scientists to make changes in specific genes far faster and in a much more precise way than ever before. The technology has two components - a homing device to a specific section of DNA (guide-RNA) and enzymatic ‘scissors’ that cut DNA (Cas9 nuclease). In the cell nucleus, the guide-RNA sequence directs the Cas9 nuclease to cause double-stranded breaks in the target DNA sequence. By harnessing the cell’s own DNA-repair apparatus, the gene being targeted can be altered either by deleting it, adding nucleotides to it or by turning its activity on or off. In contrast to previous genome-editing techniques, such as zinc-finger nucleases and TALENs, CRISPR is easier to handle in the laboratory.
Dr. Mene Pangalos, Executive Vice President, Innovative Medicines & Early Development, AstraZeneca, said: “CRISPR is a simple yet powerful tool that enables us to manipulate genes of potential importance in disease pathways and examine the impact of these modifications in a highly precise way. By combining the great science from our labs with these world-renowned academic and industry partners, we will be able to integrate this ground-breaking technology into our research and help accelerate the discovery of novel treatments for patients.”
Jag hoppas att vi inte säljer Nac till AZ utan licenserar ut det till mycket högt pris för att sedan kunna licensiera till flera andra BP om resultaten är så övertygande som det verkar. Skulle AZ köpa lösningen på leverans inom genterapi hoppas jag att vi inte säljer Nac för mindre än 100 miljarder. Låter kanske galet men potentiella intjäningen är hissnande om levereringsgåtan nu är löst.
Takk for langt, spennende og informativt innlegg @Temlor!
Dette som er uthevet her er så til de grader godt å lese, igjen og igjen!
Her har vi en fundamental utfordring for CRISPR/Cas9-teknologien oppsummert. Sammenholder vi dette med det PCIB har full kontroll på, og som PW gjennomgår stort sett på hver presentasjon, nemlig endosomal escape generelt;
og FimaNAc spesielt;
og
så ser vi teknologier som tilsynelatende passer til hverandre som hånd i hanske.
Legg til uheldige bieffekter ved bruk av andre leveringsmetoder (physical delivery methods (e.g. microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles)), og at dette er et av de virkelig store forskningsområdene for tiden, med vanvittig markedsstørrelse, økende fremover, i tillegg til samarbeidet med AZ - key player på området - der samarbeidet skal finne sitt neste steg i løpet av de neste 5 ukene, så ser vi at PCIB, og vi som er aksjonærer, har en utrolig spennende tid foran oss.
Da jeg gikk inn i PCIB var det helt klart FimaChem som var årsaken, med sine sterke og lovende resultater innen Gallegangskreft. Alt annet var icing on the cake. Innen 5 uker kan icingen nå vise seg å bli lagt på før kaken er ferdig. Det blir 5 utrolig spennende uker fremover, og vi sitter på fremste rad!
The future is imminent!
Edit: La inn denne lenken: http://pcibiotech.no/wp-content/uploads/2014/01/Kan-man-levere-medisiner-med-lys.pdf
